Campy Lob Act Ere Vibrio Ra

Risk assessment
of Campylobacter spp.
in broiler chickens
FAO
FOOD AND
NUTRITION
PAPER
and Vibrio spp. in seafood

Food
and
Agriculture
Organization
of
the
United
Report of a Joint FAO/WHO Expert

Consultation
Nations
Bangkok, Thailand
5-9 August 2002
WORLD
HEALTH
ORGANIZATION
FOOD
SAFETY
CONSULTATIONS
Risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood
ii
ACKNOWLEDGEMENTS
The Food and Agriculture Organization of the United Nations and the World Health Organization
would like to express their appreciation to the expert drafting groups (see Annex 2) for the time
and effort which they dedicated to the preparation of thorough and extensive technical documents
on risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafoods. The
deliberations of this expert consultation were based on these documents.
iii
iv
Contents
INTRODUCTION........................................................................................................................................1
BACKGROUND...........................................................................................................................................1
OBJECTIVES OF THE CONSULTATION .............................................................................................2
SUMMARY OF THE GENERAL DISCUSSIONS...................................................................................3
RISK ASSESSMENT OF THERMOPHILIC CAMPYLOBACTER SPP. IN BROILER CHICKENS4
5.1.1 Summary of the Risk Assessment.......................................................................................................4

5.1.2 Introduction.......................................................................................................................................4
5.1.3 Scope .................................................................................................................................................4
5.1.4 Approach...........................................................................................................................................5
5.1.5 Key findings.....................................................................................................................................10
5.1.6 Risk assessment and developing countries ......................................................................................11
5.1.7 Limitations and caveats...................................................................................................................12
5.1.8 Gaps in the data ..............................................................................................................................12
5.2 REVIEW OF THE RISK ASSESSMENT ..........................................................................................................13
5.2.1 Introduction: ...................................................................................................................................13
5.2.2 Hazard identification ......................................................................................................................13
5.2.3 Exposure assessment:......................................................................................................................13
5.2.4 Hazard characterization .................................................................................................................15
5.2.5 Risk characterization ......................................................................................................................16
5.2.6 Risk assessment and developing countries ......................................................................................16
5.2.7 Data deficiencies.............................................................................................................................16
5.3 UTILITY AND APPLICABILITY....................................................................................................................17
5.4 RESPONSE TO THE SPECIFIC RISK MANAGEMENT QUESTIONS POSED BY THE CODEX COMMITTEE ON FOOD
HYGIENE ............................................................................................................................................................18
5.5 CONCLUSIONS AND RECOMMENDATIONS .................................................................................................18
6
RISK ASSESSMENT OF VIBRIO SPP. IN SEAFOOD.........................................................................19
6.1 SUMMARY OF THE RISK ASSESSMENTS ......................................................................................................19
6.1.1 Introduction: Vibrio spp. in seafood ...............................................................................................19
6.1.2 Scope ...............................................................................................................................................20
6.1.3 Vibrio parahaemolyticus in raw oysters consumed in Japan, New Zealand, Canada, Australia and
the United States of America ........................................................................................................................20
6.1.4 Vibrio parahaemolyticus in bloody clams .......................................................................................28
6.1.5 Vibrio parahaemolyticus in finfish ..................................................................................................31
6.1.6 Vibrio vulnificus in raw oysters......................................................................................................33
6.1.7 Choleragenic Vibrio cholerae O1 and O139 in warm-water shrimps for export............................37
6.2 REVIEW OF THE RISK ASSESSMENTS.........................................................................................................41
6.2.1 Introduction: Vibrio spp. in seafood ...............................................................................................41
6.2.2 Vibrio parahaemolyticus in raw oysters consumed in Japan, New Zealand, Canada, Australia and
the United States of America oysters............................................................................................................42
6.2.3 Vibrio parahaemolyticus in bloody clams .......................................................................................42
6.2.4 Vibrio parahaemolyticus in finfish ..................................................................................................42
6.2.5 Vibrio vulnificus in raw oysters......................................................................................................43
6.2.6 Choleragenic Vibrio cholerae O1 and O139 in warm-water shrimp for export .............................43
6.3 UTILITY AND APPLICABILITY ....................................................................................................................43

6.4 RESPONSE TO THE SPECIFIC QUESTIONS POSED BY THE CODEX COMMITTEE ON FISH AND FISHERY
PRODUCTS .........................................................................................................................................................44
6.5 RESPONSE TO THE NEEDS OF THE CODEX COMMITTEE ON FOOD HYGIENE ...............................................46
6.6 CONCLUSIONS AND RECOMMENDATIONS ..................................................................................................46
7
CONCLUSIONS.........................................................................................................................................47
8
RECOMMENDATIONS ...........................................................................................................................47
ANNEX 1: LIST OF PARTICIPANTS.............................................................................................................49

ANNEX 2: LIST OF WORKING DOCUMENTS ...........................................................................................51
vi
List of Abbreviations
CAC
CCFFP
CCFH
CFU
CT
ctx
FAO
FDA-VPRA
g
GHP
GMP
h
ISSC
MPN
PCR
ppt
RAP
TDH
tdh
TLH
TRH
trh
USFDA
WHO
Codex Alimentarius Commission

Codex Committee on Fish and Fishery Products
Codex Committee on Food Hygiene
colony forming unit
cholera toxin
cholera toxin gene
Food and Agriculture Organization of the United Nations
The United States Food and Drug Administration draft risk
assessment on the "Public Health Impact of Vibrio
parahaemolyticus in Raw Molluscan Shellfish"
gram(s)
Good Hygienic Practices
Good Manufacturing Practices
hour(s)
Interstate Seafood Sanitation Conference
Most Probable Number
Polymerase Chain Reaction
parts per thousand
FAO Regional Office for Asia and the Pacific
Thermostable direct haemolysin
Thermostable direct haemolysin gene
Thermolabile haemolysin
TDH-related haemolysin
tdh-related haemolysin gene
United States Food and Drug Administration
World Health Organization
vii
viii
1 Introduction
The Food and Agriculture Organization of the United Nations (FAO) and the World Health
Organization (WHO) convened an expert consultation on Risk assessment of Campylobacter spp. in
broiler chickens and Vibrio spp. in seafood in the FAO Regional Office for Asia and the Pacific
(RAP), Bangkok, Thailand on 5 - 9 August 2002. The list of participants is presented in Annex 1.
Mr Dong Qingsong, FAO Deputy Regional Representative for Asia and the Pacific and
Officer-in-charge, RAP, opened the meeting on behalf of the two sponsoring organizations. In
welcoming the participants Mr Qingsong noted the increasing significance of microbiological hazards
in relation to food safety. He noted that international trade had amplified the opportunity for these
hazards to be disseminated from the original point of production to locations thousands of miles away,
thereby permitting such food safety hazards to impact on public health and trade in more than one
country. Mr Qingsong observed that this underlined the need to first consider microbiological hazards
at the international level and provide the means by which they can then be addressed at regional and
national levels. He highlighted the commitment of FAO and WHO to provide a neutral international
forum to consider new approaches to achieving food safety, and in particular to address
microbiological risk assessment.
As the meeting was convened in Asia, Mr Qingsong also highlighted the importance of the
work on microbiological risk assessment for this region. He noted that seafood and poultry are
important products in both domestic and international trade and provide a valuable contribution to the
economy of the region. He also noted the concerns for public health raised by the microbiological
hazards associated with these foods. Therefore, in conclusion he requested the expert consultation to
consider, in particular, the practical application of their work so that the output would be of value to a
range of end users and to countries in different stages of development.
The consultation appointed Dr Serv Notermans (the Netherlands) as chairperson of the
consultation and Ms Dorothy-Jean McCoubrey (New Zealand) as rapporteur. Dr Notermans also
served as chairperson of the campylobacter working group and Prof. Diane Newell (United Kingdom)
served as rapporteur. Dr Mark Tamplin (United States of America) served as chairperson of the
working group on vibrio and Dr Ron Lee (United Kingdom) as rapporteur of that group.
2 Background
Risk assessment of microbiological hazards in foods has been identified as a priority area of
work for the Codex Alimentarius Commission (CAC). At its 32nd session the Codex Committee on
Food Hygiene (CCFH) identified a list of pathogen-commodity combinations for which it requires
expert risk assessment advice. In response, FAO and WHO jointly launched a programme of work
with the objective of providing expert advice on risk assessment of microbiological hazards in foods
to their member countries and to the CAC.
Dr. Hajime Toyofuku, WHO, and Dr. Sarah Cahill, FAO provided participants with an
overview of the joint FAO/WHO activities on microbiological risk assessment. In their presentation,
they also highlighted the objectives and expected outcomes of the current meeting.
The FAO/WHO programme of activities on microbiological risk assessment aims to serve
two customers - the CAC and the FAO and WHO member countries. The CAC, and in particular the
CCFH, needs sound scientific advice as a basis for the development of guidelines and
recommendations as well as the answers to specific risk management questions on certain pathogencommodity combinations. Member countries on the other hand need specific risk assessment tools to
use in conducting their own assessments and, if possible, some modules that can be directly applied in
a national risk assessment.
To implement this programme of work, FAO and WHO are convening a series of joint expert
consultations. To date three expert consultations have been held. The first of these was held on 17 - 21
July 20001 and the second on 30 April - 4 May 20012. Both of these expert consultations addressed
risk assessment of Salmonella spp. in broiler chickens and eggs and Listeria monocytogenes in readyto-eat foods. In March 2001, FAO and WHO initiated risk assessment work on the two pathogencommodity combinations being considered in this expert consultation. Two ad hoc expert drafting
groups were established to examine the available relevant information on Campylobacter spp. in
broiler chickens and Vibrio spp. in seafood and prepare documentation on both the exposure
assessment and hazard characterization steps of the risk assessment. These documents were reviewed
and evaluated by a joint expert consultation convened on 23 - 27 July 20013.
In October 2001, the report of that expert consultation was delivered to CCFH. Additional
risk management guidance was sought from the Committee in relation to the finalization of the risk
assessments. In response to this, the Committee established risk management drafting groups to
consider the approach it could take towards providing guidance on managing problems associated
with Campylobacter spp. in broiler chickens and Vibrio spp. in seafood. A limited amount of
feedback was received; therefore the risk assessment drafting groups, FAO and WHO met early in
2002 to consider how to complete the risk assessment work. Both risk assessments have been
progressed towards completion, although to varying extents. The risk assessment documents that have
been developed to date were reviewed and evaluated by the expert consultation.
The purpose of this report is to present the summary of the draft documents on risk
assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood as well as the
discussions and recommendations of the expert consultation.
3 Objectives of the consultation

The expert consultation examined the information provided by the expert drafting groups on
risk assessment of Campylobacter spp. in broiler chickens and Vibrio spp. in seafood with the
following objectives;
To critically review the documents prepared by the ad hoc expert drafting groups giving
particular attention to;
the risk characterization component and the manner in which the risk assessment
outputs were generated;
the assumptions made in the risk assessment;
the associated uncertainty and variability;
the potential application of the work.
To provide scientific advice to FAO and WHO member countries on the risk assessment of
Vibrio spp. in seafood and Campylobacter spp. in broiler chickens based on the available
documentation and the discussions during the expert consultation.
To respond to the risk management needs of Codex.
FAO. 2000. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods, FAO
Headquarters, Rome, Italy, 17 - 20 July 2000. FAO Food and Nutrition Paper 71.
http://www.fao.org/es/ESN/food/risk_mra_campylobacter_en.stm / http://www.who.int/fsf/Micro/index.htm
2
FAO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods: Risk
characterization of Salmonella spp. in eggs and broiler chickens and Listeria monocytogenes in ready-to-eat foods, FAO
Headquarters, Rome, Italy, 30 April - 4 May 2001. FAO Food and Nutrition Paper 72.
http://www.fao.org/es/ESN/food/risk_mra_salmonella_en.stm / http://www.who.int/fsf/Micro/index.htm
3
WHO. 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
Foods; Hazard identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens
and Vibrio spp. in seafood. WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
4 Summary of the general discussions

The drafting groups presented overviews of the risk assessment documents on Campylobacter
spp. in broiler chickens and Vibrio spp. in seafood to the expert consultation. A summary of the draft
risk assessments and the discussions of the expert consultation are given in sections five and six of
this report. However, a number of issues relevant to risk assessment in general that were addressed are
summarized below.
It was anticipated that the campylobacter and vibrio risk assessments will prove useful for
decision makers such as Codex committees, food industries, food control authorities, regulators and
other stake holders. It was not the task of the risk assessment groups to undertake risk profiling or
determine acceptable risk levels this is the role of risk managers. However, the outputs of this work
should provide tools and information useful to both risk managers and risk assessors.
It was not possible at this stage to complete full quantitative risk assessments for all
pathogens, inclusive of full validation exercises. This was due to the lack of extensive quantitative
data being available, the uncertainties that exist and the need to make assumptions e.g. that all strains
of Campylobacter spp. are pathogenic. However, it was noted that even preliminary quantitative and
good qualitative risk assessments would be very useful in decision making. The risk assessment of
Vibrio parahaemolyticus in bloody clams provides an example of how readily accessible data can be
collated into the risk assessment framework to give valuable information that could be used towards
improving public health.
The models on Campylobacter spp. and Vibrio spp provided examples of how risk assessment
can be applied to pathogen-commodity combinations. However, it is important that countries do not
just transfer any conclusions from this risk assessment work to make risk management decisions
without giving due consideration to the effect of different species, environments and populations in
their own country.
The challenge will be to show the world, including developing countries, the advantages of
undertaking risk assessment. Training, complete with easy to understand guidelines on how to apply
the risk assessments was considered extremely important. The training and guidelines material should
be written in simple language that is applicable to different audiences and countries with different
levels of development.
There is concern by many countries that not having a full risk assessments will create trade
barriers. Assistance may need to be given to many developing countries to generate and collect data
and undertake modelling exercises so that such concerns can be addressed.
The consultation formed two working groups to address the risk assessment documentation
on Campylobacter spp. and Vibrio spp. respectively. The composition of the two working groups is
outlined in the table below.
Campylobacter spp. in broiler chickens
Independent experts
John Cowden
Heriberto Fernndez
Geoffrey C. Mead
Diane G. Newell
Serv Notermans
Sasitorn Kanarat
Paul Brett Vanderlinde
Expert members of the drafting group

Bjarke Christensen
Aamir Fazil
Emma Hartnett
Anna Lammerding
Maarten Nauta
Greg Paoli
Members of the secretariat

Sarah Cahill
Karen Huleback
Jeronimas Maskeliunas
Hajime Toyofuku
Vibrio spp. in seafood

Independent experts
Nourredine Bouchriti
Jean-Michel Fournier
Ron Lee
Carlos A. Lima dos Santos
Dorothy Jean McCoubrey
Marianne Miliotis
Mitsuaki Nishibuchi
Pensri Rodma
Mark Tamplin
Expert members of the drafting group

Anders Dalsgaard
Angelo DePaola
I. Karunasager
Thomas McMeekin
Ken Osaka
John Sumner
Mark Walderhaug
Members of the secretariat

Lahsen Ababouch
Peter Karim BenEmbarek
Maria de Lourdes Costarrica
5 Risk Assessment of thermophilic Campylobacter spp. in

broiler chickens
5.1.1 Summary of the Risk Assessment
5.1.2 Introduction
An understanding of thermophilic Campylobacter spp., and specifically Campylobacter jejuni
in broiler chickens is important from both public health and international trade perspectives. In order
to achieve such an understanding, evaluation of this pathogen-commodity combination by quantitative
risk assessment methodology was undertaken.
The first steps of this process, hazard identification, hazard characterization and the
conceptual model for exposure assessment were presented at an expert consultation in July 2001, and
summarized in the report of that consultation4. The exposure assessment model described the
production-to-consumption pathway for fresh and frozen broiler chicken carcasses prepared and
consumed in the home.
The final step, risk characterization, integrates the exposure and dose-response information in
order to attempt to quantify the human-health risk attributable to pathogenic thermophilic
Campylobacter spp. in broiler chickens. The risk assessment model and results were presented to the
present expert consultation in the working document MRA 02/01. Recommendations for
modifications and/or areas for further development of the exposure assessment model, arising from
the 2001 consultation were taken into consideration and, where possible, were incorporated during
preparation of the MRA 02/01 report. Recommendations, that were not incorporated, are noted in the
exposure assessment summary section below.
5.1.3 Scope
The purpose of the work was to develop a risk assessment that attempts to understand how the
incidence of human campylobacteriosis is influenced by various factors during chicken rearing,
processing, distribution, retail storage, consumer handling, meal preparation and finally consumption.
A benefit of this approach is that it enables consideration of the broadest range of intervention
4
strategies. The risk characterization estimates the probability of illness per serving of chicken
associated with the presence of thermophilic Campylobacter spp. on fresh and frozen whole broiler
chicken carcasses with the skin intact and which are cooked in the domestic kitchen for immediate
consumption. A schematic representation of the risk assessment is shown in Figure 5.1.
Dose
Response
Prevalence
Farm
&
Transport
Slaughter
&
Processing
Preparation
&
Consumption
RISK
Concentration
FIGURE 5.1: Schematic representation of the risk assessment of Campylobacter spp. in broiler
chickens.
5.1.4 Approach
5.1.4.1 Hazard identification
Thermophilic Campylobacter spp. are a leading cause of zoonotic enteric illness in most
developed countries (Friedman et al., 20005). Human cases are usually caused by C. jejuni, and to a
lesser extent, by Campylobacter coli. Information on the burden of human campylobacteriosis in
developing countries is more limited (Oberhelman & Taylor, 20006). Nevertheless, it is reported that
asymptomatic infection with C. jejuni and C. coli is frequent in adults. In children, under the age of
two, C. jejuni, C. coli and other Campylobacter spp. are all associated with enteric disease.
Campylobacter spp. may be transferred to humans by direct contact with contaminated
animals or animal carcasses or indirectly through the ingestion of contaminated food or water. The
principal reservoirs of thermophilic Campylobacter spp. are the alimentary tracts of wild and
domesticated mammals and birds. Campylobacter spp. are commonly found in poultry, cattle, pigs,
sheep, wild animals and birds, and in dogs and cats. Foodstuffs, including, poultry, beef, pork, other
meat products, raw milk and milk products, and, less frequently, fish and fish products, mussels and
fresh vegetables can also be contaminated (Jacobs-Reitsma, 20007). Findings of analytical
epidemiological studies are conflicting. Some have identified handling raw poultry and eating poultry
products as important risk factors for sporadic campylobacteriosis (Friedman, et al., 20005), however
others have found that contact in the home with such products is protective (Adak et al., 19958).
Friedman, C., Neimann, J., Wegener, H. & Tauxe, R. 2000. Epidemiology of Campylobacter jejuni infections in the United
States and other industrialized nations. In Campylobacter 2nd Edition, pp. 121-138. Edited by I. Nachamkin & M. J. Blaser.
Washington DC: ASM Press.
6
Oberhelman, R. & Taylor, D. 2000. Campylobacter infections in developing countries. In Campylobacter 2nd Edition, pp.
139-154. Edited by I. Nachamkin & M. Blaser. Washington DC: ASM Press.
7
Jacobs-Reitsma, W. 2000. Campylobacter in the food supply. In Campylobacter 2nd Ed., pp. 467-481. Edited by I.
Nachamkin & M. J. Blaser. Washington DC: ASM Press.
8
Adak, G. K., Cowden, J. M., Nicholas, S. & Evans, H. S. 1995. The Public Health Laboratory Service national case-control
study of primary indigenous sporadic cases of campylobacter infection. Epidemiology and Infection 115, 15-22.
Information for the hazard identification section was compiled from published literature and
from unpublished data submitted to FAO and WHO by public health agencies and other interested
parties.
5.1.4.2 Hazard characterization
Hazard characterization provides a description of the public health outcomes following
infection, including sequelae, pathogen characteristics influencing the organism's ability to elicit
infection and illness, host characteristics that influence the acquisition of infection, and food-related
factors that may affect the survival of C. jejuni in the human gastrointestinal tract. A dose-response
model was derived that mathematically describes the relationship between the numbers of organisms
that might be present in a food and consumed (the dose), and the human health outcome (the
response). In order to achieve this, human feeding trial data (Black et al., 19889) for two strains of
C jejuni were pooled and used to derive estimates for the probability of infection (colonization of the
gastrointestinal track without overt symptoms) as well as the probability of illness once infected.
Campylobacteriosis predominantly occurs as sporadic cases (Friedman, et al., 200010), and hence
dose-response data from outbreak investigations are essentially non-existent.
The probability of infection upon ingestion of a dose of C. jejuni was estimated with the
caveat that the data are from a feeding study involving healthy volunteers, and using a milk matrix
and a limited number of campylobacter strains. Whether the probability of infection and/or illness are
different for immune individuals or those individuals with an increased susceptibility to illness (e.g.
immunosuppressed, very young, etc.) compared to the volunteers could not be determined from the
feeding trial data. The probability of illness following infection was also estimated based upon these
investigations, and assumed to be dose-independent. Again, the impact of other factors, such as
susceptibility, on the probability of illness cannot be quantified due to a lack of adequate
epidemiological data and resolution to this level. The progression of the illness to more serious
outcomes and the development of some sequelae can be crudely estimated from the approximate
proportions reported in the literature, but these were not included in the present work. For the
purposes of this model it was assumed that C. coli has the same properties as C. jejuni.
5.1.4.3 Exposure assessment
The exposure model describes the stages Rearing and Transport, Slaughter and Processing,
and Preparation and Consumption (Figure 5.1). This modular approach estimates the prevalence and
concentration of thermophilic Campylobacter spp., and the changes in these associated with each
stage of processing, and refrigerated and frozen storage and consumer handling of the product.
Routes contributing to initial introduction of Campylobacter spp. into a poultry flock11 on the
farm have been identified in epidemiological studies. However, these remain poorly understood and
the phenomenon may be multi-factorial, and hence, colonization was modelled based on an
unspecified route of introduction. However, an epidemiology module has been created as a
supplement to the model, which allows evaluation of interventions aimed at changing the frequency
with which risk factors occur. As such risk factors are usually interdependent and may be country
specific, incorporation of this module into the current risk model was not appropriate: however, it
does illustrate how such information may be used in a specific situation.
The effects of cooling and freezing, and adding chlorine during water chilling, are evaluated
in the model. Effects of other processing treatments, e.g. lactic acid and irradiation, on reduction of
Campylobacter spp. contamination on poultry carcasses were not explicitly evaluated as the
quantitative data are lacking: however, if the level of reduction as a consequence of any type of
9
Black, R. E., Levine, M. M., Clements, M. L., Hughes, T. P. & Blaser, M. J. 1988. Experimental Campylobacter jejuni
infection in humans. Journal of Infectious Diseases 157, 472-9.
10
Friedman, C., Neimann, J., Wegener, H. & Tauxe, R. 2000. Epidemiology of Campylobacter jejuni infections in the
United States and other industrialized nations. In Campylobacter 2nd Edition, pp. 121-138. Edited by I. Nachamkin & M. J.
Blaser. Washington DC: ASM Press.
11
A flock is a group of hens of similar age that are managed and housed together.
processing modification, can be quantified for any treatment, then the impact on risk can be readily
evaluated within the risk model.
The home preparation module considered two routes of ingestion of viable organisms: via
consumption of contaminated, undercooked chicken meat, and via cross-contamination, from
contaminated raw poultry onto another food that is eaten without further cooking, or directly from the
hands (Figure 5.2). As a result of recommendations from the previous expert consultation12 the
protected areas cooking approach and the drip-fluid model approach for cross-contamination were
selected. The "protected areas" approach to modelling the cooking step considers that campylobacter
cells may be located in parts of the carcass that reach cooking temperature more slowly than other
locations. The drip-fluid model for cross-contamination relates to the spread of campylobacter cells
via drip-fluid from raw carcasses.
Raw Chicken
Cross contamination
Under cooking
Exposure
FIGURE 5.2: Schematic representation of the home preparation module.
5.1.4.4 Risk characterization
The risk characterization step integrates the information collected during the hazard
identification, hazard characterization, and exposure assessment steps to arrive at estimates of adverse
events that may arise due to consumption of chicken (Figure 5.1). This step links the probability and
magnitude of exposure to campylobacter associated with consumption of chicken to adverse outcomes
that might occur. The resulting risk is expressed as individual risk or the risk per serving of chicken.
Although this model does not address risk to a specific population, data on amounts of chicken
consumed can be incorporated into the model to arrive at estimates of the population-based risk.
The current model is unable to provide a central estimate of risk due to virtually unbounded
uncertainty13 on two key components of the model, namely the impact of undercooking and the
impact of cross-contamination. Since these processes are the ultimate determinant of the final
exposure of the consumer, the unbounded and unresolvable nature of the uncertainty undermines the
establishment of a central estimate of consumer risk. In addition, the model has not yet been applied
12
13
As the upper and lower boundaries of the values describing the impact of undercooking and cross-contamination are
unknown the distribution of possible values is potentially infinite and therefore unbounded.
to explore the upper and lower bound estimates of risk that are suggested by the uncertainty.
However, the model still provides a useful means of studying potential exposure pathways and how
these contribute to the risk of illness posed by campylobacter associated with broiler chickens.
The risk characterization involves the following elements:
1) A baseline model
A baseline model was explored in detail, to clarify the input-output relations between the
different modules of the model and also to explore the credibility of the model. The baseline model
was defined as the processing of fresh carcasses from both positive and negative flocks (with an
overall flock prevalence of 80%), which are air chilled at the end of slaughter.
The model indicated that the external campylobacter load per chicken increased during
transport and evisceration, and decreased at the other processing steps studied, with an overall
reduction of the mean load from farm to fork of about 4 to 5 logs (Figure 5.3). The prevalence of
campylobacter-contaminated chickens from positive flocks appears to drop from 100% of live birds to
20% of chicken meat servings (Figure 5.4). For negative flocks, prevalence increases during transport,
defeathering and evisceration, indicating the effect of cross-contamination during processing.
Prevalence later drops to a value of about 3% of servings at the moment of consumption.
The correlation between the input and output of the models of the different processing steps
has been graphed to illustrate the variability in load per chicken along the process. These plots
indicate that the load on a chicken at the stages before evisceration appears to be a bad predictor for
the final load, and thus for the risk per serving of that particular chicken carcass.
2) Scenario analysis
In a scenario analysis, the effects of alterations to the baseline are studied by changing one or
two of the model parameters. By doing so the impact of uncertainty in parameter estimates can be
explored, and the potential of risk mitigation strategies can be evaluated.
Eight alternative scenarios were explored, to illustrate the ability of the model to investigate
the effect of changing specific model inputs. Each individual scenario involved a specific change in
one parameter, as follows:
Mitigation related scenarios
1. Between flock14 prevalence (80% prevalence reduced to 50%)
2. Within flock prevalence (100% prevalence reduced to 10%)
3. Scalding (hard scald (56-58C for 2-2.5 minutes)and soft scald (50-52 C for up to
3.5 minutes))
Alternate model assumption scenarios
4. Shorter freezing storage time range (1 day to 6 weeks storage time compared to 1 day
to 1 week storage time)
5. Undercooking (5C lower temperature in coolest spot in the chicken during cooking)
6. Defeathering (an increase the magnitude of cross-contamination)

7. Contamination during water chilling without chlorine (a decrease of the
contamination added)
8. Contamination during water chilling with chlorine (decrease of the contamination
added)
The largest effects were observed with lower undercooking temperature, which led to an increase in
risk and lower within flock prevalence, which led to a decrease in risk.
14
Between flock refers to different groups of hens (i.e. different flocks).
10
8
6
4
2
0
se
do
r
s t e fri
or g.
ag
e
in
g
as
h
w
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r
at
io
n
rin
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ev
is
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a
th
e
ng
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di
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-2
FIGURE 5.3: Numbers of campylobacter cells per carcass during processing of fresh air-chilled
carcasses, from both positive and negative flocks. (Negative flocks get contaminated during
transport). Both the mean of the logs and the log of the means are given. (The differences between
these is as a result of the skewness15 of the distribution of values and the fact that 'zero'- values cannot
be incorporated in calculations of the mean of logs (which is therefore only about the positive
log mean of positive flocks
carcasses)). ____________
mean log of positive flocks
log mean of negative flocks
mean log of negative flocks
do
se
tra
ns
po
rt
sc
ald
ing
de
fe
at
he
rin
g
ev
isc
er
at
ion
wa
sh
ing
re
f ri
g.
sto
ra
ge
1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0.0
fa
rm
Prevalence
----------______ ______
x
------x------
FIGURE 5.4: The prevalence of campylobacter on fresh air-chilled broiler chicken carcasses coming
from flocks that were positive and negative for campylobacter at the farm level.
Flocks positive for campylobacter at the farm
Flocks negative for campylobacter at the farm
15
Skewness is a statistical term that refers to the lobsideness of a distribution.
To further detail the model analysis, and to determine the effect of introducing different
mitigations, five series of simulations were run. Using the baseline model settings one of following
parameters was changed in each series of simulations:
flock prevalence;
campylobacter load on carcasses after transport;
campylobacter load in caeca;
campylobacter load in both caeca and on carcasses after transport;
campylobacter load on carcasses after evisceration.
A linear relationship between flock prevalence and probability of illness was found. Thus a
two-fold reduction in flock prevalence would result in a corresponding two-fold reduction in the
probability of illness. This effect is mainly caused by a reduction in the number of campylobacterpositive meals. Both external contamination and colonization need to be reduced concurrently to
achieve a substantial impact on risk. This is due to the potential for large numbers of campylobacter
cells contaminating the carcass as a result of damage to the viscera at evisceration, thus undermining
any benefits achieved in reducing the external numbers at an earlier processing step. Use of the model
has identified two key ways of achieving this reduction: 1) reducing colonization early enough to
impact external contamination and 2) intervening post-evisceration to reduce carcass contamination.
5.1.5 Key findings

1. There is a lack of systematic and fundamental investigation into the key processes throughout the
production-to-consumption continuum that may lead to human infection as a result of chicken
consumption. This was evidenced by the extensive knowledge acquired and data gaps identified
during this risk assessment.
2. Quantifying and characterizing uncertainty, although often recommended, needs to be recognized
as a task that can be unfeasible, and relatively unrewarding, in situations where the model
contains:
a. many uncertain parameters (lack of data and information to inform the parameters of the
model),
b. a large amount of uncertainty (lack of data and information to inform the mathematical
description of how the system works, including causal relationships and dependencies), or
c. a risk assessment simulation model that is complex (constraints in computing ability in
order to perform the quantification).
In the case of this risk assessment, all three of these properties apply. Although there were
numerous quantifiable uncertainties, it was believed that model uncertainties (point b, above)
would dominate the total uncertainty. The extent of the model uncertainties was such that the
magnitude of the quantifiable uncertainties would be rendered irrelevant.
3. It was difficult to model cross-contamination in the home as a result of the lack of a clear
understanding of the pathways and lack of data quantifying the magnitude and frequency of crosscontamination events. Further improvement of this module and validation may be extremely
difficult given the complexity of cross-contamination, the many possible pathways by which it
can occur, and the variability in the behaviour of individuals in the kitchen.
4. Based on thermal inactivation calculations, it was difficult to reconcile the assumed importance of
undercooking as a cause of human exposure to campylobacter, if the contamination of broiler
carcasses with campylobacter is on the external surface of the carcass (or very close to the
surface). Resolution of this inconsistency requires the allocation of some amount of
contamination to sub-surface sites within the carcass where the temperature increases much more
slowly. While it is possible to demonstrate that campylobacter will, on occasion, be found in such
places, it is very difficult to quantify the frequency and extent of this particular mode of
contamination.
10
5. Overall campylobacter concentration on chicken carcasses decreased through processing, with

temporary increases occurring during transport and evisceration.
6. The prevalence of campylobacter-positive carcasses from negative flocks increased up to and
including evisceration and decreased at later stages. This decrease after evisceration was also
found for positive flocks, depending upon the method of chilling.
7. The campylobacter load on a chicken at the stages before evisceration was identified as a bad
predictor for the final load, and thus for the risk per serving of that particular chicken.
8. Assuming that cooking performance is independent of the chicken being fresh or frozen, frozen
chicken posed a lower risk via consumption than fresh chicken.
9. The washing-off effect associated with water chilling translated to water-chilled chickens posing a
lower risk than air-chilled chickens. However, there was uncertainty associated with the degree of
cross-contamination that occurs in the chill tank during water chilling that would have an impact
on this comparison and may be affected by the addition of chlorine to the chill water.
10. Undercooking was estimated to have a higher risk than cross-contamination using one set of
assumptions. The cooking and cross-contamination modules are based on plausible theoretical
constructs, but knowledge and data related to these two pathways are essentially unavailable.
Since the set of assumptions on which this comparison relied was only one, of many, plausible
sets, analysis of this component of the model remains inconclusive.
11. Unlike many other mitigation scenarios, there was very little uncertainty that reduction of the
between-flock prevalence of campylobacter would reduce any associated public health risk. A
linear relationship was found to exist between flock prevalence and probability of illness, i.e. a
two-fold reduction in flock prevalence would result in a corresponding two-fold reduction in the
probability of illness.
12. In order to meaningfully reduce the bacterial load on processed carcasses, interventions would be
required to address the bacterial load both internally and externally, since efforts directed at only
one of these contamination reservoirs can be readily undermined by high levels of contamination
from the other.
5.1.6 Risk assessment and developing countries

The current risk assessment document also addressed the issue of whether it is possible for
developing countries to apply the concepts and use the components from this model to conduct their
own quantitative risk assessment. The relative complexity of the current model was recognized. It is
relevant to poultry produced and processed under conditions similar to those described in the risk
assessment and so, in particular, may be applicable to the large-scale production and processing
facilities in developing countries. However, many of the exposure elements in developing countries the consumption patterns, slaughter processes and farming practices - may be quite different from
those described here, thus limiting the applicability of the risk assessment. Furthermore, there are few,
if any, data on exposure routes, risk factors and human illness associated with campylobacter in
developing countries. Thus, the possibility of performing a national quantitative microbial risk
assessment may require a capacity that does not currently exist in many developing countries. There
are steps that developing countries can take to aid future risk assessment efforts. A knowledge of risk
management activities, which initiate and facilitate the risk assessment process, are important. The
"Guidelines for incorporating quantitative risk assessment in the development of microbiological food
hygiene standards, guidelines and related texts"16 currently being elaborated by FAO and WHO
include guidance on preliminary risk management activities which are critical in structuring the risk
16
The process of elaborating FAO/WHO "Guidelines for incorporating quantitative risk assessment in the development of
microbiological food hygiene standards, guidelines and related texts" began at a consultation convened in Kiel, Germany on
18 - 22 March 2002. The draft guidelines are currently being finalized and will be available on the FAO and WHO webpages
at the end of 2002. http://www.fao.org/es/ESN/food/risk_mra_riskmanagement_en.stm / http://www.who.int/fsf/Micro/index.htm
11
assessment process. Data collection, one of the most important related activities, received particular
attention and focus was given to the main tasks that need to be undertaken.
5.1.7 Limitations and caveats

The expert consultation identified features of the assessment that have an impact on the
acceptability of the model and the appropriateness of using this model. The model was developed so
as not to be representative of any specific country or region. Yet many of the model inputs were based
on data and processing practices in one country. Prior to using the model to predict risk for a specific
country, data, that is representative of that country, should be used to determine model inputs.
A hypothetical baseline scenario was considered in order to evaluate the relative merit of
control strategies for campylobacter in broiler chickens. This scenario consists of a compromise of
input assumptions derived from a range of countries. The findings from the baseline scenario should
not be used to draw inferences about a particular nation or region. To make specific inferences it
would be necessary to collect inputs that were directly relevant to the particular population of interest.
Although the uncertainty associated with several parameters in the consumption portion of the
risk assessment was accounted for, a full analysis of statistical and model uncertainty was not
performed. This is explained in section 5.1.4. (under 2).
The data available for generation of the dose-response curve was limited to one feeding trial
study (see section 5.1.3.2). An alteration to our current understanding of the dose-response
relationship, which may occur if for example additional dose-response information became available,
would result in changes in the risk estimates generated by the model.
5.1.8 Gaps in the data

In the course of undertaking this risk assessment, it was found that appropriate data was not
always available and this limited the extent to which the risk assessment could be completed. The
main data gaps that were identified are outlined below.
Exposure assessment: On-farm
Survey data on the prevalence of campylobacter-positive flocks for slaughter, that
includes information on sample size, test methods etc.
Data on the probability of contamination of birds during transport.
Studies on the dynamics of within-flock transmission of campylobacter.
Data on the routes of campylobacter infection in broilers.
Exposure assessment: Processing
Prevalence and enumeration data for campylobacter on carcasses before and after various
processing steps such as scalding, defeathering, evisceration, washing and chilling.
Prevalence and enumeration data for campylobacter on carcasses comparing various
methods of chilling (e.g. air chilling, water chilling, water chilling with chlorine).
Prevalence and enumeration data for campylobacter on carcasses comparing different
scalding temperatures or alternate scalding configurations (e.g. multi-tank scalding
systems).
Data describing the actual cross-contamination between positive and negative flocks and
within flocks during the different slaughter processes.
Exposure assessment: Post-processing and consumer handling
Additional data on the cooking of chicken that addresses areas of the chicken where
campylobacter may be protected from heat.
Survey data and direct observational data on consumer practices in preparation and
handling of chicken that especially detail the frequency and degree that transfer of
campylobacter and subsequent ingestion of campylobacter could occur.
Hazard Characterization
12
Data on strain variability regarding virulence/pathogenicity and survival during

processing.
Studies on the mechanisms of infectivity, virulence/pathogenicity of campylobacter in the
human host.
Quantitative information about infection and illness rates at low doses of C. jejuni, and
also at a range of doses of different strains of C. jejuni and strains of C. coli, from 100 to
109 organisms.
Complete epidemiological data from outbreak studies including enumeration of
thermophilic campylobacter in suspected food items or in drinking water, numbers of
people exposed, attack rates, and demographics of those exposed, particularly
immunocompromised population groups and children under the age of five.
Data describing the impact of and longevity of acquired immunity resulting from recent
exposure to thermophilic campylobacter.
5.2 Review of the Risk Assessment

The expert consultation reviewed the document entitled Preliminary Report - A Draft Risk
Assessment of Campylobacter spp. in Broiler Chickens (MRA02/01) and the presentation of
additional data by the drafting group. It acknowledged the extensive work undertaken by the risk
assessment drafting group and reviewed the components of the risk assessment document, the
outcome of which is summarized below.
5.2.1 Introduction:
The expert consultation highlighted the importance of including in the introduction of the
final risk assessment document a succinct, clear description of the history of this exercise: by whom it
was initiated; what reports have been produced, and when. The objectives of the risk assessment
should be more clearly stated, with reference to who suggested them. Objectives which have been
specifically excluded should also be listed. It would be useful to state whether each objective has been
achieved and if not, why not. This would facilitate a better understanding of the work and why it was
developed in such a manner. Descriptions of methods and approaches should not appear in this
section.
A concluding paragraph suggesting the steps to be taken following the successful conclusion
of this initiative would be beneficial to the end-users, while also stressing that no risk assessment
model is complete as long as data gaps exist.
5.2.2 Hazard identification

The final risk assessment document will require an up-to-date review of hazard identification
associated with Campylobacter spp. and campylobacteriosis in humans and animals.
5.2.3 Exposure assessment:

5.2.3.1 Campylobacter in poultry on the farm and during transportation:
The main features of this module are:
A model of a single point source of infection in the broiler house. This model is
adaptable to investigate alternative sources should data become available.
A tool for estimating the probability that a random bird will be campylobacterpositive at the point of slaughter.
A model of the external contamination of birds during catching and transportation.
This part of the model assumes that:
13
All strains have the same colonization potential and persistency in the chicken gut:
with the current level of knowledge this is the only assumption that can be made, but
it may be untrue.
There are only three transmission scenarios.
Birds stay in their social clusters during harvesting. However, it is known that birds
move during harvesting so the social clusters are unlikely to be maintained.
Data deficiencies and recommendations for model improvements
It is evident that the reduction or elimination of campylobacter colonization in the flock is
extremely important. At the farm level there are limited strategies to achieve this aim. Approaches
include preventing flock exposure by biosecurity or reducing bird susceptibility to colonization by
measures such as vaccination or competitive exclusion treatment (Newell & Wagenaar, 200017). The
latter approaches are not yet available commercially and, therefore, biosecurity is the only strategy
currently feasible. A module to assess the relative importance of sources of colonization would be
extremely useful to risk managers, and such a module has been initiated. However, it was considered
by the risk assessment drafting group that there were at this time insufficient data on flock infection
sources for the use of such a module.
The part of the model dealing with the effect of catchers contaminating the exterior of birds
could, with some modification, be adapted to model the effects of thinning (the early removal of a
proportion of the birds) which is considered a major source of broiler colonization.
Using data on between-flock and within-flock prevalence of campylobacter the probability of
any random bird being campylobacter-positive can be estimated. The model also demonstrates that
the probability of colonization is dependant on age which is consistent with available data (Newell
and Wagenaar, 200017). Although the model can indicate the effect of various generic sources on
transmission it cannot, at this time, allow an assessment of the sources of exposure to provide targeted
strategies for intervention.
5.2.3.2 Processing
The main features of this module are:
A model which mimics the changes in level of contamination after scalding,
defeathering, evisceration and chilling.
A provision in the model to differentiate the effects of air and water chilling and the
use of super-chlorination.
An estimation of the prevalence of contaminated products
A provision to model the changes in level of contamination after storage at 4 oC or
frozen at -20 oC.
Campylobacter do not grow outside the host gut. This is consistent with most
scientific evidence (Jacobs-Reitsma, 200018).
There are a finite number of sites of cross-contamination within a plant. There are
however many more sites of potential cross-contamination within a processing plant
other than scalding, defeathering and evisceration but these would appear to include
the most important sources (Mead 198919).
The poultry production industry has substantially improved hygiene control over the last 30
years. Many of the changes may have effects on the data entered into the model, for example the
17
Newell, D. G. & Wagenaar, J. A. 2000. Poultry infections and their control at the farm level. In Campylobacter, 2nd Ed.,
pp. 497-509. Edited by I. Nachamkin & M. J. Blaser. Washington DC: ASM Press.
18
Jacobs-Reitsma, W. 2000. Campylobacter in the food supply. In Campylobacter 2nd Ed., pp. 467-481. Edited by I.
Nachamkin & M. J. Blaser. Washington DC: ASM Press.
19
Mead G.C. 1989. Hygiene problems and control of process contamination. In: Processing of poultry (ed Mead G.C.)
Chapman Hall, London.pp 183-220.
14
introduction of multistage scalding and newer evisceration machinery which separates the viscera
from the carcass. If there are data or evidence that such changes have a significant effect on
campylobacter contamination of carcasses then appropriate changes to the model should be
considered.
Overall, the spread of contamination from the gut contents of positive birds will have the
greatest effect on the level of contamination on external bird surfaces. However, as the proportion of
negative flocks increases, the importance of cross-contamination becomes more apparent. Data is now
being collected for the level of contamination on carcasses from campylobacter-negative flocks. This
data should be available in the near future, and the expert consultation recommended that at that time
it should, if possible, be incorporated into the model.
The model should take account of the published and unpublished data showing that organisms
attached to the carcass surface are resistant to environmental effects. (Notermans and Kemplemaker,
197520; G. Mead, personal communication, 2002).
Only freezing and chilled storage were considered in the post-processing part of this module.
Recent anecdotal data indicates that modified atmosphere packaging has an impact on levels of
campylobacter contamination on poultry products. The expert consultation recommended that when
this data comes into the public domain it should, if possible, be incorporated into this module.
There is increasing evidence for variation in the survival of campylobacter strains during
processing (Newell et al., 200121). This may have a considerable effect on the model especially if the
ability of Campylobacter spp. to survive is associated with strain virulence. At this time there are no
simple methods for assessing the ability to survive but models for this property are available and data
is currently being accumulated. Once this data becomes available the expert consultation
recommended that it should, if possible, be incorporated into this module.
5.2.3.3 Consumer handling and cooking
The main features of this part of the model are:
Estimates of the proportion of loosely attached campylobacter cells associated with
poultry.
Estimates of the concentration of campylobacter cells in the drip-fluid from a random
wet chilled carcass.
A model of the effect of temperature, time of exposure and location at a protected site
on campylobacter survival.
10-20% of campylobacter cells are located at protected sites. The expert consultation
considered this assumption to be unexpectedly high.
More information is required on consumer practices in domestic kitchens. Information is
needed on routes and modes of transmission of campylobacter within the kitchen. Factors affecting
campylobacter survival in the kitchen environment are also required.
5.2.4 Hazard characterization

Due to the lack of new data on the dose-response relationship for campylobacter, no further
progress can been made in this area. Similarly there is currently a paucity of data on the proportion of
campylobacter strains in poultry which are virulent in humans and to which humans are susceptible.
20
Notermans S. and Kamplemaker E.H. 1975. Heat destruction of some bacterial strains attached to broiler skin. British
Poultry Science, 16: 351-361
21
Newell, D. G., Shreeve, J. E., Toszeghy, M., Domingue, G., Bull, S., Humphrey, T. & Mead, G. 2001. Changes in the
carriage of campylobacter strains by poultry carcasses during processing in abattoirs. Applied and Environmental
Microbiology. 67, 2636-40.
15
5.2.5 Risk characterization

As highlighted in section 5.1.3.4 there is a lot of uncertainty associated with the current
model, primarily due to the lack of information on the impact of undercooking and crosscontamination on exposure of the consumer to campylobacter. The expert consultation could not
foresee that this uncertainty would be significantly reduced in the near future. However, despite this
situation, the expert consultation noted that the model could be applied to explore the sensitivity of the
risk estimates to a broad range of plausible alternate models and parameters. In addition, as much
contextual information as possible on the uncertainty and its implication should be provided for risk
managers and also researchers in this area so that consideration can be given as to how this
uncertainty can be reduced.
The model remains very useful to study the biological and systematic plausibility of alternate
hypotheses regarding these exposure pathways and will contribute to the understanding of the risk
posed through these two pathways.
The expert consultation noted that for certain combinations of model parameters a 10-fold
reduction in the probability of infection from a single serving had a dramatic effect on the probability
of illness (about 4-fold). However, only preliminary results were available as work was ongoing to
refine this section. In doing so the expert consultation recommended that the effect combined
mitigations (e.g. vaccination and freezing) also be evaluated using the model.
5.2.6 Risk assessment and developing countries

It was noted that the problem of campylobacteriosis in developing countries, at a national
public health level, may be considerably different from that in developed countries (Oberhelman and
Taylor, 200022). Epidemiological features of campylobacteriosis are distinct and suggest more
frequent exposures from various sources. There is substantial and increasing evidence that immunity
as a consequence of repeated exposure plays an important role in protection against
campylobacteriosis (Cawthraw et al., 200023) and this may be particularly relevant in developing
countries (Newell & Nachamkin, 199224). It was recommended that such countries take this factor
into account when applying the model.
One of the most important issues for developing countries is whether the application of the
risk assessment model is appropriate at all. The model may be usefully adapted for the exporting
poultry industry in such countries, as it is anticipated that such commercial operations will be similar
to those in developed countries. However, for domestic public health purposes, the current risk
assessment model would need considerable adaptation. In particular, chicken processing practices
may be less standardized and more variable. Moreover, food habits are likely to vary and the immune
status of the population is expected to be different.
5.2.7 Data deficiencies

There are numerous data gaps which have precluded the development of a complete risk
model. Therefore, the deliberate omission of uncertainty analysis in the model was accepted by the
expert consultation.
It was recognized that, for further development of the model, new data are clearly needed and
are also required to facilitate selection of appropriate interventions measures by risk managers.
22
Oberhelman, R. & Taylor, D. 2000. Campylobacter infections in developing countries. In Campylobacter 2nd Edition, pp.
139-154. Edited by I. Nachamkin & M. Blaser. Washington DC: ASM Press.
23
Cawthraw, S. A., Lind, L., Kaijser, B. & Newell, D. G. 2000. Antibodies, directed towards Campylobacter jejuni antigens,
in sera from poultry abattoir workers. Clinical and Experimental Immunology 122, 55-60.
24
Newell, D. & Nachamkin, I. 1992. Immune responses directed against Campylobacter jejuni. In Campylobacter jejuni:
Current staus and future trends, pp. 201-206. Edited by I. Nachamkin, M. Blaser & L. Tompkins. Washington DC: ASM
Press.
16
The expert consultation identified the following research priorities, however, it is critical to
note that research priorities will vary depending on the specific risk management question the risk
assessment is being used to address.
A. Exposure assessment
Information about the influence of strain-specific variation on campylobacter survival on
poultry meat.
Information on consumer and retail practices in relation to the risk of transfer of
campylobacter from poultry products to kitchen surfaces and other foods.
Data on the routes of campylobacter colonization of broilers at the farm level so that farm
interventions can be appropriately targeted.
B. Hazard characterization
Additional data on dose-response.
Data on strain variability in relation to virulence and pathogenicity.
5.3 Utility and Applicability

The expert consultation recognized this risk assessment as a resource that can be used by
many parties including national authorities. The current document provides a framework for
undertaking risk assessment of thermophilic Campylobacter spp. in broiler chickens, a modelling
approach that can be adapted to various situations and a unique source of data and other relevant
information. While it does not overcome the need to establish a risk assessment team at the national
level it will significantly reduce the workload of such a team and the time they require to carry out a
risk assessment. This exercise has also led to the identification of the types of data that are required
for risk assessment. While, it was noted that the collection of such data will depend on the purpose for
which the risk assessment is being undertaken, the risk assessment nonetheless highlights a number of
areas where data generation activities may need to be focussed.
Furthermore, the model in its current form can be used by risk managers to help them make
decisions on the suitability of specific interventions. Given information on the efficacy of an
intervention, the relative reduction in the number of possible cases of illness can be assessed. More
specific issues in relation to the utility of this work are elaborated on below.
Applicability of the model to different production systems
The model described in the risk assessment has been developed to estimate the risk of
thermophilic campylobacter from broiler chickens produced under a specified production system. The
model is an example of how a risk assessment could be undertaken and details mathematical models
that might be used to describe production practices. In order for the model to be used in different
countries it must be modified to reflect prevailing practices.
Utility of the model to risk assessors
The model contains all the elements necessary to enable risk modellers to perform risk
assessments for their own production systems. The modular approach used enables modellers to
utilize various components of the model individually or collectively. When finally presented the
model should be structured in a clear and concise way. This is important if the model is to be used as
a tool to aid modellers in conducting risk assessments.
Utility of the model to Risk Managers
Communication is a critical element in ensuring the risk assessment model is of use to risk
managers. The model should not be used by risk managers without the aid and advice of competent
risk assessors, who can explain the assumptions and uncertainties associated with the model. For
example in the current model cooking and cross-contamination are not modelled on real data,
because such data are not yet available. The expert consultation noted the limitations placed on the
utility of the model by the absence of evidence and lack of consensus among experts on the relative
importance of cross-contamination.
17
5.4 Response to the specific risk management questions posed by

the Codex committee on food hygiene
As noted by the previous expert consultation on Campylobacter spp. in broiler chickens25 the
risk management questions posed by the CCFH were not tailored to the particular problem of
campylobacter in chicken.
At a meeting of a CCFH drafting group26, established by the committee to develop a
discussion paper on risk management strategies for campylobacter in poultry, it was considered that
the provision of guidance to the risk managers on the relative efficacy of mitigation strategies would
be a useful outcome of the risk assessment. Given this, it was decided that interventions at various
points in the overall process would be investigated rather than the investigation of any specific
mitigation strategy.
Five scenarios were selected, one dealing with the effect of changing the flock prevalence and
four others looking at the effect of reducing campylobacter load, either on the exterior of the chickens
at slaughter or in the chickens gut before slaughter. The outcome of these scenarios are presented in
the risk characterization and key findings sections (5.1.4.4 & 5.1.5).
Applying the campylobacter risk assessment to particular regions or areas will require the
collection and input of data specific for local conditions. Similarly the risk assessment model may
need adaptation. The risk assessment model should be used by a risk analysis team including risk
assessors. The needs of risk managers must be considered as must the limitations imposed by
economic, political, consumer and stakeholder priorities.
5.5 Conclusions and Recommendations

The risk assessment model utilizes a modular approach that is applicable to the entire poultry
supply chain. It is flexible to use and capable of dealing with a range of issues relating to
campylobacter contamination and its control in poultry from production-to-consumption. The exercise
described in the report has covered both the conceptual development of the model and the evaluation
of data needed to demonstrate its value. For practical application, however, the model would need to
be modified, adapted or even redeveloped to suit the differing circumstances of individual users. The
model takes no account of uncertainty because data are lacking on this aspect. In relation to
developing countries, the model is particularly relevant to conditions of intensive production and
processing that occur where poultry meat is exported. While it could also be adapted to the free range
village chicken that is a feature of many developing countries, the expert consultation was not
aware of any evidence on the risk to public health from this type of bird.
The expert consultation recognized the uncertainties surrounding the relative importance of
undercooking and cross-contamination in the kitchen but, from epidemiological evidence, were of the
opinion that cross-contamination was the more important factor.
The expert consultation made a number of recommendations aimed at improving the
transparency and utility of the risk assessment document:
The layout of the document should be clarified.
The major assumptions made in the study should be listed.
The benefits to the poultry industry from applying the model should be fully explained.
A simplified description of the model should be made available to risk managers.
It should be emphasized that the practical application of the model, in whatever form, will require
training and guidance of putative users, particularly in developing countries.
25
WHO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods; Hazard
identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens and Vibrio spp. in
seafood, WHO Headquarters, Geneva, Switzerland 213 - 27 July 2001. WHO 2001.
26
Report of the thirty fourth session of the Codex Committee on Food Hygiene, Bangkok, Thailand, 8 - 13 October 2001
ALINORM 03/13 para. 77. http://www.codexalimentarius.net/reports.asp
18
Possible intervention measures should be given in the report to illustrate actions that could be
taken by risk managers.
Technical details given in the background information on production and processing need to be
updated.
6 Risk assessment of Vibrio spp. in seafood

6.1 Summary of the risk assessments
6.1.1 Introduction: Vibrio spp. in seafood
Vibrio spp. are Gram-negative, facultatively anaerobic motile curved rod-shaped bacteria with
a single polar flagellum. The genus contains at least twelve species pathogenic to humans, ten of
which can cause food-borne illness (Table 6.1). The majority of food-borne illness is caused by
Vibrio cholerae, Vibrio parahaemolyticus or Vibrio vulnificus (Oliver and Kaper, 199727; Dalsgaard,
199828). Most countries have guidelines for detecting V. parahaemolyticus and V. cholerae O1 and
O139 in seafood, whereas few have guidelines for V. vulnificus. Accordingly, routine microbiological
analysis of seafood includes testing for V. parahaemolyticus and V. cholerae O1 and O139, but
seldom for V. vulnificus.
Some species are primarily associated with gastrointestinal illness (V. cholerae and
V. parahaemolyticus) while others can cause non-intestinal illness, such as septicaemia (V. vulnificus).
In tropical and temperate regions, disease-causing species of Vibrio occur naturally in marine, coastal
and estuarine (brackish) environments and are most abundant in estuaries. Pathogenic vibrios, in
particular V. cholerae, can also be recovered from freshwater reaches of estuaries (Desmarchelier,
199729), where it can also be introduced by faecal contamination. The occurrence of these bacteria
does not generally correlate with numbers of faecal coliforms and depuration of shellfish may not
reduce their numbers. However, a positive correlation between faecal contamination and levels of
V. cholerae may be found in areas experiencing cholera outbreaks. A positive correlation between
water temperature and the numbers of vibrios has also been shown in several parts of the world.
Further, according to data from the United States of America and Denmark, there is a positive
correlation between water temperature and both the number of human pathogenic vibrios isolated and
the number of reported human infections. This correlation is particularly striking for
V. parahaemolyticus and V. vulnificus (Dalsgaard et al., 199630).
The objective of the work was to undertake a risk assessment of Vibrio spp. in seafood
products that have the most impact on public health and/or international trade. Three species,
V. parahaemolyticus, V. vulnificus and choleragenic V. cholerae (toxigenic V. cholerae O1 and O139
that may cause cholera) were identified as being responsible for most illnesses caused by Vibrio spp.
The approach taken was to quantify those illnesses caused by Vibrio spp. in different countries
following the consumption of a range of seafoods and this report documents the results of that
approach.
27
Oliver, J. D., and Kaper, J.B. 1997. Vibrio Species. In M. P. Doyle, L. R. Beuchat, and T. J. Montville, eds. Food
Microbiology: Fundamentals and Frontiers, p228-264. Washington, D.C., ASM Press.
28
Dalsgaard, A. 1998. The occurrence of human pathogenic Vibrio spp. and Salmonella in aquaculture. International
Journal of Food Science and Technology, 33: 127-138.
29
Desmarchelier, P.M. 1997. Pathogenic Vibrios. In A.D. Hocking, G. Arnold, I. Jenson, K. Newton and P. Sutherland, eds.
Foodborne Microorganisms of Public Health Significance 5th Edition, p 285 -312. North Sydney, Australian Institute of
Food Science and Technology Inc.
30
Dalsgaard, A. Mller, N.F., Brin, B., Hoei, L. and Larsen, J.L. 1996. Chemical manifestation and epidemiology of Vibrio
vulnificus in Denmark (summer 1999). European Journal of Clinical Microbiology and Infectious diseases 15. 227 - 232.
19
6.1.2 Scope
The risk assessment work was undertaken on the following pathogen-commodity
combinations:
Vibrio parahaemolyticus in raw oysters consumed in Japan, New Zealand, Canada,
Australia and the United States of America.
Vibrio parahaemolyticus in finfish consumed raw.
Vibrio parahaemolyticus in bloody clams consumed in Thailand.
Vibrio vulnificus in raw oysters consumed in the United States of America.
Choleragenic Vibrio cholerae in warm-water shrimp in international trade.
TABLE 6.1: Vibrio spp. which cause, or are associated with, human infections (after Dalsgaard,
199831)
Occurrence in human clinical specimens*
Intestinal
V. cholerae O1 and O139
V. cholerae non-O1/non-O139
V. parahaemolyticus
V. fluvialis
V. furnissii
V. hollisae
V. mimicus
V. metschnikovii
V. vulnificus**
V. alginolyticus
V. carchariae
V. cincinnatiensis
V. damsela
++++
++
++++
++
++
++
++
+
+
-
Non-intestinal
+
++
+
+
+
+++
++
+
+
+
*The symbol (+) refers to the relative frequency of each organism in clinical specimens and (-) indicated that the organism
was not found
**The ability of V. vulnificus to cause gastro-intestinal disease remains to be confirmed
6.1.3 Vibrio parahaemolyticus in raw oysters consumed in Japan,

New Zealand, Canada, Australia and the United States of
America
6.1.3.1 Introduction
FAO and WHO aim to make optimal use of existing risk assessments in their MRA activities.
As there have been large outbreaks of illness due to V. parahaemolyticus in North America following
consumption of raw oysters, the United States Food and Drug Administration (USFDA)
commissioned a quantitative risk assessment on the "Public Health Impact of Vibrio
parahaemolyticus in Raw Molluscan Shellfish" (FDA-VPRA), one output of which was the
31
Dalsgaard, A. 1998. The occurrence of human pathogenic Vibrio spp. and Salmonella in aquaculture. International
Journal of Food Science and Technology, 33: 127-138.
20
development of a risk model. A critical component of the model was water temperature. Since high
water temperatures are a factor in several countries with significant oyster industries, FAO and WHO
decided to undertake a risk assessment on consumption of raw oysters in a number of different
countries. As well as generating an estimate of the number of annual illnesses, a further aim was to
assess the potential of the model developed in the United States of America to predict oyster-borne
V. parahaemolyticus illness from oysters grown in different regions and using different production
systems.
6.1.3.2
Scope
The risk assessment covers consumption of raw oysters in five countries: New Zealand,
Japan, Canada, Australia and the United States of America.
V. parahaemolyticus has been recognized as a major cause of seafood-borne gastroenteritis in
Japan (Twedt, 198932; Ministry of Health, Labour and Welfare, Japan, 200033) and other Asian
countries. By contrast, in most countries outside of Asia, the reported incidence appears to be low,
perhaps reflecting a different mode of seafood consumption. Gastroenteritis caused by this organism
is almost exclusively associated with seafood consumed raw or inadequately cooked, or contaminated
after cooking. In the United States of America, prior to 1997, illness was most commonly associated
with crabs, oysters, shrimp and lobster (Twedt, 198932; Oliver and Kaper, 199734). Four
V. parahaemolyticus outbreaks associated with the consumption of raw oysters were reported in the
United States of America in 1997 and 1998 (DePaola et al., 200035). A new V. parahaemolyticus
clone of O3:K6 serotype emerged in Calcutta in 1996. It has spread throughout Asia and to the United
States of America elevating the status of V. parahaemolyticus to pandemic (Matsumoto et al., 200036).
In Australia, in 1990 and 1992, there were two outbreaks of gastroenteritis caused by
V. parahaemolyticus in chilled, cooked shrimps imported from Indonesia (Kraa, 199537) and there was
also a death in 1992 associated with the consumption of oysters.
This section focuses on evaluating the nature of adverse health effects associated with
V. parahaemolyticus in seafood and how to quantitatively assess the relationship between the
magnitude of the food-borne exposure and the likelihood of adverse effects occurring. It included the
elaboration of a dose-response curve. Infection by V. parahaemolyticus is characterized by an acute
gastroenteritis. Therefore, the end-point of the dose-response curve was defined as gastroenteritis.
A review of the literature was undertaken to identify and characterize the infectivity and
genetic factors of V. parahaemolyticus, which has both pathogenic and non-pathogenic forms based
on the presence of specific virulence genes: tdh (thermostable direct haemolysin gene) and trh (TDHrelated haemolysin gene). Relevant factors with respect to the host and food matrix have been
identified and where data are available may be incorporated into the model.
32
Twedt, R. M. 1989. Vibrio parahaemolyticus. In M. P. Doyle, ed. Foodborne Bacterial Pathogens, p543-568. New York,
Marcel Decker, Inc.
33
Ministry of Health, Labour and Welfare, Japan 2000. Statistics of Food Poisoning Japan in 2000.
34
35
DePaola, A., C.A. Kaysner, J.C. Bowers, and D.W. Cook. 2000. Environmental investigations of Vibrio parahaemolyticus
in oysters following outbreaks in Washington, Texas, and New York (1997, 1998). Applied and Environmental
Microbiology, 66: 4649-4654.
36
Matsumoto, C., J. Okuda, M. Ishibashi, M. Iwanaga, P. Garg, T. Rammamurthy, H. Wong, A. DePaola, Y.B. Kim, M.J.
Albert, and M. Nishibuchi. 2000. Pandemic spread of an O3:K6 clone of Vibrio parahaemolyticus and emergence of related
strains evidenced by arbitrarily primed PCR and toxRS sequence analysis. Journal of Clinical Microbiology, 38: 578-585.
37
Kraa, E. 1995. Surveillance and epidemiology of food-borne illness in NSW, Australia. Food Australia, 47(9): 418- 423.
21
The determination of the dose-response relationship was based on the best available data.
Human volunteer studies were available for the construction of the dose-response curve for
V. parahaemolyticus, however, these studies characterize the dose-response relationship for
V. parahaemolyticus administered with a pH-neutralizing buffer rather than with a food matrix. The
data were analysed using curve-fitting routines to find a best fit for the Beta-Poisson dose-response
curve. Because of the limited amount of data available from human volunteer studies the resulting
dose-response relationship is uncertain. This uncertainty was accounted for by representing the doseresponse relationship in the form of a family of plausible data-derived dose-response curves
determined using resampling techniques. Figure 6.1 shows the most probable dose-response curve for
V. parahaemolyticus; however, the family of curves representing uncertainty that surrounds the curve
is not shown.
Probability of Gastroenteritis
0.1
0.01
0.001
0.0001
0.00001
0
10
12
log dose of V.parahaemolyticus

10
FIGURE 6.1: Beta-Poisson dose-response curve for V. parahaemolyticus (endpoint modelled is

gastrointestinal illness)
Beta-Poisson
Sanyal and Sen (1974)38
Aiso and Fujiwara (1963)39
Takikawa (1958)40

In the United States of America, during 1997 and 1998, there were more than 700 cases of
illness due to V. parahaemolyticus, the majority of which were associated with the consumption of
raw oysters. In two of the 1998 outbreaks a serotype of V. parahaemolyticus previously reported only
in Asia, O3:K6, emerged as a principal cause of illness for the first time. It was suggested that warmer
than usual water temperatures were responsible for the outbreaks.
Temperature profiles in the oyster industries of Japan, New Zealand, Australia, Canada and
the United States of America were obtained, together with consumption levels of oysters and bacterial
levels of V. parahaemolyticus in the oysters. The objectives were to quantify the exposure of
consumers to pathogenic V. parahaemolyticus from consumption of raw oysters in these countries.
38
Sanyal, S.C., and Sen P.C. 1974. Human volunteer study on the pathogenicity of Vibrio parahaemolyticus. In T. Fujino,
G. Sakaguchi, R. Sakazaki, and Y. Takeda. eds. International Symposium on Vibrio parahaemolyticus. p. 227-230. Tokyo,
Saikon Publishing Company.
39
Aiso K & Fujiwara K. 1963. Feeding tests of the pathogenic halophilic bacteria. Annual Research Report Institute of Food
Microbiology Chiba University, 15:34-38.
40
Takikawa I. 1958. Studies on pathogenic halophilic bacteria. Yokohama Medical Bulletin, 9:313-322.
22
The FDA-VPRA model was used as the base to accommodate data inputs from other countries. This
model incorporates all phases in the harvest - post-harvest consumption continuum in three modules
(Figures 6.2-6.4).
Figure 6.2 shows a conceptual model for the harvest module. Water temperature is the driving
input with regard to the initial numbers of V. parahaemolyticus in oysters. In the way the analysis is
constructed regional and seasonal temperature variations allow for a multi-year analysis that can
account for long-term temperature trends. Water salinity is shown in dotted lines to indicate that for
some model applications salinity may be another important input.
R egional, seasonal & yearly variation
W ater salinity
W ater tem perature
total Vp/g
pathogenic V p/g
FIGURE 6.2:Harvest module for exposure assessment of V. parahaemolyticus in oysters.

(Vp = Vibrio parahaemolyticus)
T im e to refrigeration
V p/g at harvest
A ir tem p erature
V p/g at 1st refrigeration
C oold ow n tim e
V p/g at cooldow n
S torage tim e
V p/g at con su m ption
FIGURE 6.3: Post-harvest module for exposure assessment of V. parahaemolyticus in oysters.

(Vp = pathogenic Vibrio parahaemolyticus)
Figure 6.3 shows the conceptual model for post-harvesting practices. The post-harvest module
determines the role of post-harvest processing and handling on the numbers of pathogenic
23
V. parahaemolyticus at consumption. The bubble denoting "V.p/g at harvest" is the output of the
harvest model shown in Figure 6.2. Inputs on the time the oysters are out of the water and the air
temperature are used to predict growth of V. parahaemolyticus in the oysters. Growth continues as the
oysters are cooled but at a different rate. V. parahaemolyticus levels decrease during storage and the
storage time is therefore an input time that affects V. parahaemolyticus numbers.
Figure 6.4 represents the consumption module. The bubble denoting "path Vp/g (numbers) at
consumption" is the output of the post-harvest module. This number is multiplied by the number of
oysters per serving and the weight of the oysters to yield the ingested dose. This ingested dose is used
in the dose-response to calculate the risk of illness associated with the consumption of one oyster
meal.
V.p./g
(numbers) at
consumption
# oysters
per serving
Ingested
dose
Weight per
oyster (g)
RISK OF
ILLNESS
FIGURE 6.4: Consumption module for exposure assessment of V. parahaemolyticus in oysters

(Vp = pathogenic Vibrio parahaemolyticus)

The data from the five countries were analysed for incorporation into the risk assessment
model. The risk assessment model was modified to allow for a monthly analysis of data from Japan,
Australia, and New Zealand. The analysis for Canada and the United States of America was done on a
seasonal basis. Using the Japanese data only one simulation, consisting of 100 000 iterations, was
undertaken, as multiple year temperature data were not available. Thirteen simulations, consisting of
10 000 iterations were undertaken for Australia reflecting the availability of 13 years of data. As only
one year's data were available for New Zealand, one simulation, consisting of 100 000 iterations, was
undertaken. For Canada 1 000 simulations, consisting of 10 000 iterations, based on United States of
America Pacific Northwest data, was undertaken. The analysis for the United States of America
consisted of 10 000 iterations for seasons in four regions.
6.1.3.7 Key findings
Introduction
The complete data sets required to test the applicability of the model to harvesting waters of
countries other than the United States of America were not available. In particular tdh+ and trh+ data
were lacking and in such cases United States of America data was used as a surrogate to allow for
testing of the model.
24
Japan
Based on the available data set41, the preliminary predictions of illness are shown in Table
6.2. The model predicted low levels of illness for November to April. The model was not run for the
months of May to October as oysters for raw consumption are not harvested during this period42.
It was difficult to compare this with epidemiological data for V. parahaemolyticus-related
oyster illnesses in Japan for a number of reasons. The Japanese surveillance system focuses mainly on
outbreaks of food-borne disease and therefore the number of laboratory confirmed reported illnesses
may not include sporadic cases or diffuse outbreaks and the extent of under-reporting is not known
(K Osaka, personal communication, 2002). In addition the food source of the illness may not always
be identified. However, in cases where oysters have been identified as the food source causing illness,
large variability in the annual number of V. parahaemolyticus-related oyster illnesses has been noted
over the last five years43. It is also worth noting that the model estimation is based on data (e.g. air and
water temperature, salinity) available from only one of the major harvesting areas and therefore does
not necessarily capture the situation in the different oyster growing areas in Japan.
TABLE 6.2: Preliminary predictions of V. parahaemolyticus illness in Japan associated with oyster
consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
4
Second quarter
(Apr-Jun)
1
(April only42)
Third quarter
(Jul-Sep)
042
Fourth quarter
(Oct-Dec)
19642
(Nov-Dec
only)
Total
201
Australia
Based on the available data set, the preliminary predictions of illness are shown in Table 6.3.
The model predicted more illnesses than the number of reported cases (J. Sumner, personal
communication, 2002). The application of United States of America surrogate data to a different
species of oyster, specifically the Sydney rock oyster, may have a role in the overestimation of risk.
TABLE 6.3: Preliminary predictions of V. parahaemolyticus illness in Australia associated with
oyster consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
157
Second quarter
(Apr-Jun)
28
Third quarter
(Jul-Sep)
10
Fourth quarter
(Oct-Dec)
33
Total
228
New Zealand
The model predicted more illnesses than the number of reported cases (D.J. McCoubrey,
personal communication, 2002) (Table 6.4). As extensive use of surrogate data from the United States
41
Japanese water temperature is available at http://www.hiroins-net.ne.jp/suisansc/suion.html; Air temperature data came

from the Japanese Meteorological Agency; Ogawa, H. Tokunou, H., Kishmoto, T., Fukuda, S., Umemura, K. & Takata, M.
(1989) Ecology of V. parahaemolyticus in Hiroshima Bay. Hiroshima. Journal of Veterinary Medicine No. 4. (in Japanese);
Consumption data came from "Family income and expenditure survey (2000) (The Japan Institute of Labour) and "The
national nutrition survey" (1995) (Japanese Ministry of Health and Welfare)
42
Oysters for raw consumption are not harvested in Japan from May to October in the area where the data was collected
because Microbiological criteria (MPN of coliform group in harvesting seawater, the total plate count numbers, MPN of
coliform groups and MPN of total Vibrio parahaemolyticus in oyster ) are exceeding the standards set by the Ministry of
Health, Labour and Welfare.(Personal communication, Ken Osaka)
43
Anonymous. 1999. Report of Food Poisoning , Japan 1997-2001, Ministry of Health, Labour and Welfare, Japan.
25
of America was necessary as inputs for some of the parameter required to run the model, the true risk
may be much lower than that predicted.
TABLE 6.4: Preliminary predictions of V. parahaemolyticus illness in New Zealand associated with
oyster consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
13
Second quarter
(Apr-Jun)
17
Third quarter
(Jul-Sep)
0
Fourth quarter
(Oct-Dec)
5
Total
35
Canada
The preliminary results (Table 6.5) indicate that model predicted cases of illness that are
relatively close to the number of reported cases 4445). The proximity of the Canadian harvesting waters
to one of the regions of the United States of America that was modelled allows greater confidence in
these predictions. It should be noted that the model did not consider the mitigation to cool oysters
immediately after harvest that was introduced in the Canadian oyster industry in 2000, as the data
used was collected prior to the implementation of this measure.
TABLE 6.5: Preliminary predictions of V. parahaemolyticus illness in Canada associated with oyster
consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
0
Second quarter
(Apr-Jun)
1
Third quarter
(Jul-Sep)
7
Fourth quarter
(Oct-Dec)
0
Total
8
United States of America

The predicted numbers of illness for the United States of America as shown in Table 6.6. In
this case the dose-response relationship was adjusted to take into account the estimation that the actual
number of cases of V. parahaemolyticus illness in the United States of America exceeds the reported
number of cases by a factor of 20 to 1 (Mead et al, 199946). However, it was acknowledged that the
predicted number of illnesses associated with oyster consumption is probably still an overestimation
as the study of Mead et al (1999)46 that estimated the degree of under-reporting used statistics on the
annual incidence of V. parahaemolyticus illness and not only those for which oyster was the vehicle
of transmission. Evidence for validation of the model comes from the observed agreement between
model predictions of V. parahaemolyticus numbers with observed harvesting and retail numbers of
V. parahaemolyticus.
Table 6.5: Preliminary predictions of V. parahaemolyticus illness in the United States of

America associated with oyster consumption
Number of
predicted illnesses
First quarter
(Jan-Mar)
40
Second quarter
(Apr-Jun)
1587
44
Third quarter
(Jul-Sep)
3881
Fourth quarter
(Oct-Dec)
376
Total
5884
Cato, J.C. 1998. Economic values associated with seafood safety and implementation of seafood, Hazard Analysis Critical
Control Point (HACCP) programmes. FAO Fisheries Technical Paper. No. 381. Rome, FAO. 1998.
45
Anonymous, 1997, Canada Communicable Disease Report - Volume 23-19, October 1, 1997.
46
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625.
26
6.1.3.8 Limitations and Caveats

It was difficult to critically evaluate the performance of the model in harvesting waters
outside of the United States of America. In many cases the raw data on which to adapt the model to
local conditions were not available because:
The data had not been accumulated because of expense or lack of need for the data.
The data were in summary form and therefore not amenable to reanalysis.
The data were difficult to retrieve from stored printed form and to convert into electronic format.
The methodology used to generate the data in countries other than the United States of America
was not comparable to that used in generating the data that served as a basis for the establishment
of the model parameters.
Where limited data were available, judgement was needed on how to adapt this data for
incorporation into the model. However, there is currently no guidance on this issue or even whether
adaptation of data is desirable.
Validation of model predictions by epidemiological observations was complicated by the fact
that the relationship between observed and predicted illness is generally unknown. In the United
States of America the ratio of predicted to observed illness has been estimated to be 20 to 1 (Mead et
al, 199947). This relationship has not been estimated for other countries and where it may differ from
that in the United States of America.
Limited data has the effect of reducing the variance of the models prediction of risk. The
reduced variance of predictions may be misinterpreted as greater confidence in a predicted risk than a
predicted risk with wider variance that is based upon more extensive data.
The species of oyster may have a profound effect on the model and further research is needed
to develop the oyster-V. parahaemolyticus ecology knowledge base.
Accurate model predictions may require adapting the model to parameters that are critical to
harvesting areas and are different from those in the United States of America where the model was
developed. For example, salinity may be a critical element in the control of V. parahaemolyticus in
New Zealand and Australia. The model will be elaborated to test whether the addition of this
parameter can improve model predictions.
The use of surrogate data, particularly in relation to the occurrence of tdh+ and trh+ strains,
may limit the utility of the model in predicting illnesses from V. parahaemolyticus contaminated
oysters harvested from waters other than those of the United States of America. Obtaining these data
may be difficult, especially when few illnesses associated with oysters from certain harvesting areas
lead to the fact that data (required by the model) have not being collected.
6.1.3.9 Data gaps
The risk assessment identified a number of data gaps which limited in particular the
application of the model developed in the United States of America to oysters harvested in different
regions of the world. Some of the main data and knowledge gaps include:
Multi-year temperature data for seawater and air at harvesting areas.
Data for determining the temperature to V. parahaemolyticus numbers relationship in some
harvesting areas.
Characterization of harvesting activities in some areas.
Information on the role of oyster ecology in altering the model parameters.
Data for tdh+ and trh+ prevalence of the total V. parahaemolyticus in some national
harvesting waters.
Dose-response information, the lack of which results in uncertainty in the dose-response
relationship and adds substantial variance to predictions of illness.
47
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625
27
Methodology for estimating the ratio of reported to total cases of illness. Appropriate
methodology needs to be officially developed and applied in countries that wish to compare
the number of illnesses predicted by risk assessment to the number of reported & recorded
cases of illness.
Data sources that can indicate whether or not the model is succeeding. Validation of the risk
assessment model should be attempted at as many intermediate stages as is practical.
6.1.4 Vibrio parahaemolyticus in bloody clams

V. parahaemolyticus has been recognized as a major cause of food-borne gastroenteritis in
Japan and other Asian countries. However, the data available on V. parahaemolyticus and seafood,
other than oysters, that was also suitable for the quantitative risk assessment were very limited worldwide.
A small-scale study was undertaken, based on data collected in the Songkla Province of
southern Thailand. A joint Thai-Japanese team carried out the study on the prevalence and
concentration of V. parahaemolyticus in non-oyster seafood. All strains of V. parahaemolyticus and
pathogenic strains which have the tdh and / or the trh gene, and thus have the potential to produce
TDH and /or TRH, were enumerated in the data collection process. No foodborne disease surveillance
data for this area were available. However, the preliminary study showed that the strains isolated from
clinical specimens in this area were identical, in terms of serotype and molecular genetics, with the
strains isolated from the shellfish harvested in the area rather than other seafood such as fish and
shrimps. Therefore, a popular bivalve in Thailand, the bloody clam (Anadara granosa), was chosen as
the target seafood in this risk assessment. This shellfish is also traded in the Southeast Asian region.
6.1.4.2 Scope
Using state of the art techniques, the data necessary for developing the quantitative risk
assessment were collected and a model was elaborated in a developing country situation where there
was a lack of quantitative data.
V. parahaemolyticus is considered to be an important cause of seafood-borne disease in
Thailand. A survey of clinical specimens obtained from patients with diarrhoea resulted in the
isolation of 294 pathogenic strains from 317 cases that were confirmed positive for
V. parahaemolyticus (Table 6.6). Several seafood items were also tested for pathogenic strains of
V. parahaemolyticus and in this preliminary study shellfish were the most commonly contaminated
among the samples tested (Table 6.6). The profile of strains (serotype and possession of tdh / trh
gene) isolated from clinical samples were consistent with that of the strains isolated from shellfish
(Table 6.2). Therefore, shellfish were considered as an important source of V. parahaemolyticus
infection.
The dose-response model used in the hazard characterization of V. parahaemolyticus in
oysters (see section 6.1.3.4) was also used in the hazard characterization of V. parahaemolyticus in
bloody clams.
The exposure assessment was divided into four stages; harvest, retail, cooking and
consumption as shown in Figure 6.5.
Data on the prevalence and numbers of V. parahaemolyticus in clams was collected at each
step of the exposure pathway. A single lot of the clams was taken from a boat shortly after landing at
28
the harvest site. Following initial sampling (Harvest stage), the remaining clams were transported to
the local market area, which was located close to the laboratory. A sample of clams were examined at
this point to represent the Retail stage. Thereafter, the clams were maintained outside of the
laboratory for a period of time to simulate the transportation step; these were subsequently examined
in the laboratory. Typically, the clams are cooked in the home by boiling briefly (in some cases with
insufficient heating). The Cook (Boiling) stage was simulated in the laboratory and the clams were
tested thereafter. To obtain consumption data, local people were interviewed on the frequency and
quantity of bloody clams consumed.
TABLE 6.6: Results of the study on isolation of V. parahaemolyticus from seafood and the most
common strain profiles of isolates from clinical specimens and seafood
Isolation of pathogenic strains of
V. parahaemolyticus
O3:K6 tdh+, trh-
O1:K25 tdh+,
trh-
13/268 (4.4%)
0/50
0/9
0/100
8(62%)
0
0
0-
2(15%)
0
00-
294/11 375 (2.6%)
192 (65%)
22(7.5%)
Seafood samples*
Shellfish(bivalves)
Shrimp
Crab
Fish
Clinical samples**
*Samples were examined over a four year period from 1998 to 2001. During the first year of the study period pathogenic
V. parahaemolyticus were only isolated from shellfish. Therefore, during the subsequent years of the survey, efforts focussed
mainly on detection of pathogenic V. parahaemolyticus in shellfish samples.
** V. parahaemolyticus was isolated from 317 diarrhoea specimens out of a total of 11 375 samples that were examined
during a survey sporadic cases of illness with diarrhoea in 1999. Specimens came from different patients in two big hospitals
in the province. Of the 317 cases confirmed positive for V. parahaemolyticus, 294 of these were confirmed to be pathogenic
strains of V. parahaemolyticus.
Harvest
Temperature
Prevalence
Concentration
transportation
Time
Temperature
Retail
Temperature
Prevalence
Concentration
Cooking
Temperature
Prevalence
Concentration
Quantity
Frequency
transportation
Time
Temperature
Consumption
FIGURE 6.5: Schematic representation of the exposure model developed for the risk assessment of
V. parahaemolyticus in bloody clams.
29
The laboratory analysis undertaken included V. parahaemolyticus toxR gene sequencing to

identify V. parahaemolyticus and group-specific PCR (GS-PCR) to detect pandemic strains carrying
the tdh gene. The prevalence of total and tdh+ or trh+ V. parahaemolyticus strains was examined at
harvest and retail, and after cooking.
The total numbers of V. parahaemolyticus from culture and PCR methods were assumed to
have a lognormal distribution. The prevalence of total V. parahaemolyticus after boiling was
estimated using the laboratory generated data. The prevalence of tdh+ and trh+ strains that possibly
remain in clams after boiling was estimated by assuming that the ratio of the prevalence of total and
virulent strains before heating was maintained after boiling. The same assumption was made with
regard to numbers.
Comparison was made between the predicted and observed values of total
V. parahaemolyticus numbers during transportation from harvest to the retail stage, in order to
determine whether the increase in numbers could be analysed or predicted using an equation
developed in the FDA-VPRA.
Although bloody clams are a popular seafood in this region there were no available data on
their consumption. Therefore a small preliminary consumption survey was undertaken. Fourteen
people (students and workers) at the university were selected for interview because of their
accessibility. They were interviewed on how frequently they ate clams at home and how many they
ate at one meal.
The output of the exposure assessment feeds into the hazard characterization to produce the
risk characterization output. The probability of getting ill following consumption of a single serving
of clams was estimated for a defined population (i.e. people who were interviewed) by using the
dose calculated in the exposure assessment and the dose-response equation. The probability of
getting ill per year was further estimated by multiplying the frequency of clam consumption per year.
The consumption data for bloody clams were used for estimating the risk of ingesting pathogenic
strains of V. parahaemolyticus.
1. The total number of V. parahaemolyticus was estimated as 6.5 /clam, with a standard deviation of
2.2 /clam, at harvest, and 7.8 /clam, with a standard deviation of 2.0 /clam, at retail.
2. After boiling, V. parahaemolyticus was detected in only one and two out of 32 samples by PCR
and culture methods, respectively. Pathogenic strains were not isolated from any of the boiled
samples.
3. Using the data generated from culture methods, the mean probability of illness per year due to
clam consumption was estimated to be 9.18E-10 per person (approx. 1 person per 1 000 000 000
people becomes ill per year) and maximum probability was 9.34E-6 (approx. 1 person per
100 000 people becomes ill per year).
4. The observed growth rate of V. parahaemolyticus in bloody clams was found to be half the rate of
growth predicted by the FDA-VPRA V. parahaemolyticus growth rate model in oysters.
5. Although time and resources were limited and there was a lack of quantitative data, this study
indicated that, even when such obstacles exist, progress can still be made on data generation and
risk assessment modelling.
The link between human illness and consumption of bloody clams was based on detection of
strains of equivalent serotype and molecular genetics in both clinical samples and bivalve samples.
There were no data from outbreak investigations or case control studies of sporadic cases to confirm
this link or to prove that illness was indeed caused by foodborne transmission. Additional data is
required to strengthen this linkage and this should ideally be included in the risk profile that is
undertaken before the risk assessment is commissioned.
30
The results are restricted to a single food item, and the sample size may not be sufficiently
large. Therefore, the data presented in table 6.6 should be interpreted with caution. Furthermore, the
study on survival of V. parahaemolyticus from harvest to consumption was carried out for only a three
month period in one specific area in Thailand. More data are needed for other months and other areas.
Because the cooking (by boiling) module was developed based on experimental data using
fixed time / temperature values within a very limited range, scenario analysis with different time /
temperature combinations are impossible. Also the consumption survey was carried out on a small
group of people working within the same environment and therefore may not be representative of the
region as a whole.
The cross-contamination model was not applied in this risk-assessment, because of lack of
data and appropriate models for cross-contamination. Due to insufficient epidemiological data, model
validation could not be undertaken.
6.1.4.9 Gaps in the data
To improve the risk assessment, the following data will be needed.

Quantitative data on V. parahaemolyticus in bloody clams and other shellfish in various
combinations of water temperature and salinity.
The proportion of virulent strains in various shellfish, areas and seasons.
The differences in sensitivity between virulent strain and non-virulent strain to heating and other
mitigation steps.
Data from a case control study or an outbreak investigation to strengthen the linkage between
consumption of bloody clams and V. parahaemolyticus illness in humans.
6.1.5 Vibrio parahaemolyticus in finfish

V. parahaemolyticus is a leading cause of seafood-borne illness in Japan and other Asian
countries Several reports exist on the high prevalence of the organism in a variety of seafoods, in
particular finfish, lobster and shrimp. Outbreaks due to V. parahaemolyticus associated with fish and
shellfish other than oysters have been reported in some countries including the United States of
America, Thailand, China (Taiwan) and Spain. With the globalization of Japanese cuisine and the
increased practice of eating raw fish and shellfish, there is an increased possibility of
V. parahaemolyticus infection as a result of consumption of these foods. A risk assessment of
V. parahaemolyticus in finfish could provide useful information for reducing this risk.
An exposure assessment document was prepared and presented to an expert consultation48 in
2001. Although the drafting group had decided not to include this part in the final report due to the
lack of quantitative data, it was noted that, although not a complete quantitative risk assessment, it
still includes information that may be important for many countries and therefore should be recorded
and available in the public domain.
This work could currently be described as a qualitative (descriptive) risk assessment. An
effort to collect quantitative data on total V. parahaemolyticus in finfish, as well as data on virulent
strains, is not yet completed. However, should it be possible to collect the necessary quantitative data
a revised document, incorporating such data will be prepared.
48
WHO. 2001. Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in Foods: Hazard
identification, exposure assessment and hazard characterization of Campylobacter spp. in broiler chickens and Vibrio spp. in
seafood, WHO Headquarters, Geneva, Switzerland 23 - 27 July 2001. WHO 2001.
31
6.1.5.2 Scope
This work focused on describing the
V. parahaemolyticus from harvest to consumption.
possible
contamination
of
finfish
by

Published data on the prevalence and concentration of V. parahaemolyticus in finfish and
other seafood were collected and collated. Literature reviews were also conducted through Medline
and other resources on the world wide web.
The dose-response model used in the hazard characterization of V. parahaemolyticus in
oysters (see section 6.1.3.4) was also considered to be applicable in the case of V. parahaemolyticus
in finfish.
The pathway from pre-harvest to consumption was divided into four stages; pre-harvest,
harvest, post-harvest and consumption. It includes a descriptive explanation of the possible risks of
V. parahaemolyticus contamination at each stage. The possibility of proliferation / reduction of
V. parahaemolyticus in each stage was considered through a qualitative description of the data
collected.
Because insufficient data were available to bring the assessment forward, no further work was
undertaken.
1. The prevalence and numbers of V. parahaemolyticus in seawater are influenced by seawater
temperature and salinity. However, there may be other influencing factors such as plankton and
tides.
2. Many species of finfish could be contaminated with V. parahaemolyticus though the prevalence
and number of V. parahaemolyticus present vary with species. Differences in prevalence and
numbers seemed to be associated with the species and their habitat (e.g. coastal or deep-sea).
3. Coastal seawaters used at landing docks and at markets were shown to be highly contaminated
with V. parahaemolyticus. Therefore, the post-harvest stage may be of particular importance with
regard to contamination of finfish.
4. This conceptual modelling approach could be appropriate for determining the potential
effectiveness of mitigation strategies such as the use of chlorinated water and thermal processing.
5. The fluctuation of time and temperature during transportation and storage may be less important
for finfish than raw oysters as V. parahaemolyticus was shown not to proliferate significantly on
finfish samples up four hours at 25oC.
6. Washing the visceral cavity after evisceration of the intestine reduced the numbers of
V. parahaemolyticus on the fish fillet compared to eviscerated fish in which the visceral cavity
had not been washed.
7. Food preparation in the home, including the time prior to washing out the visceral cavity, was
identified as an important step in relation to cross-contamination and reducing the numbers of
V. parahaemolyticus.
This is a qualitative (descriptive) risk assessment and quantitative data on the prevalence and
concentration of V. parahaemolyticus in targeted seafoods are needed to undertake quantitative risk
assessment.
32

The lack of quantitative data prevented the completion of this risk assessment. Primarily data
in the following areas are needed.
Number and proportion of pathogenic V. parahaemolyticus cells in various species of finfish.
Frequency of consumption and quantity of raw fish consumed.
Transportation practices (time and temperature).
6.1.6 Vibrio vulnificus in raw oysters

The general approach to undertaking this assessment and many of the parameters were
adopted from the FDA-VPRA and the FAO/WHO V. parahaemolyticus risk assessment, which are the
only available quantitative risk assessments for a Vibrio spp. in raw oysters. Due to the lack of
appropriate data from outside of the United States of America for many of the model inputs this
assessment relies almost totally on data from this country. The approach for determining doseresponse used exposure and illness frequency. For this reason some elements of hazard
characterization included in the exposure assessment.
The choice of the United States of America data was intended only to provide an example on
how to apply the exposure model to a different national situation. This model could be further tested
and modified when appropriate data from other countries or situations become available.
6.1.6.2 Scope
The main objective of this risk assessment was to determine the usefulness of adapting the
FDA-VPRA model to assess the risk from V. vulnificus associated with the consumption of raw
oysters. In addition it aims to identify the most appropriate data as well as the data gaps and
limitations for modelling V. vulnificus in oysters, conduct a risk characterization of V. vulnificus in
raw oysters using available data and evaluate targeted mitigation levels aimed at reducing the risk of
V. vulnificus illness
6.1.6.3 Hazard Identification
V. vulnificus has been associated with primary septicaemia in individuals with chronic preexisting conditions, following consumption of raw bivalves. This is a serious, often fatal, disease. In
the United States of America, it carries the highest death rate of any food-borne disease agent (Mead
et al, 199949). To date, V. vulnificus seafood-associated disease has almost exclusively been
associated with oysters (Dalsgaard et al, 200150; Oliver and Kaper, 199751). In addition to the primary
septicaemia that follows ingestion, V. vulnificus is known to infect wounds of otherwise healthy
individuals, although the majority of patients with serious wound infections have an underlying
disease. Such wound infections occur most often as a result of contamination of pre-existing wounds
with seawater or after contact with fish or shellfish. V. vulnificus has in a few cases been isolated from
patients with gastrointestinal disease, however, its role as a primary cause of gastrointestinal disease
remains to be determined. Recently, cases of primary septicaemia associated with V. vulnificus
infections seem to have been related to consumption of a variety of raw seafood products in Korea
and Japan (S. Yamamoto, personal communication, 2001).
49
Mead, P.S., Slutsker, L., Dietz, V., McCaig, L.F., Bresee, J.S., Shapiro, C., Griffin, P.M. and Tauxe R.V. 1999. FoodRelated Illness and Death in the United States Emerging Infectious Diseases, 5 (5), 607-625
50
Dalsgaard, A., Hoei, L., Linkous, D. and Oliver, J.D. 2001. Vibrio vulnificus. In Y.H. Hui, M.D. Person and J. R. Gorham
(eds). Foodborne disease handbook, vol 1 Bacterial Pathogens, New York, Marcel Dekker Inc.
51
33

A schematic representation of a conceptual model of the V. vulnificus risk assessment model showing
integration of all the modules is outlined in Figure 6.6. This includes the exposure assessment
modules for harvest, post-harvest and consumption that were derived from the FDA-VPRA. The
exposure assessment examined the appropriateness of transferring inputs from the
V. parahaemolyticus risk assessment to that of V. vulnificus. Where this was not possible alternate
approaches were developed. The predicted exposure was validated with data from a survey of
V. vulnificus numbers in raw oysters at retail.
V. vulnificus can occasionally cause mild gastroenteritis in healthy individuals, but for
specific subpopulations V. vulnificus can cause a serious septicaemia that frequently leads to death in
susceptible people. Therefore, the endpoint for the dose-response curve is defined as septicaemia.
There was not adequate information to differentiate between virulent and avirulent strains of
V. vulnificus. Therefore, all V. vulnificus strains were considered to be equally pathogenic.
While data from human volunteer studies were available for the construction of dose-response
curves for V. parahaemolyticus and V. cholerae O1, no such data were available for V. vulnificus.
Therefore, an alternate approach is being attempted. The dose-response relationship can be estimated
by fitting a Beta-Poisson model using monthly data on the numbers of V. vulnificus in United States
of America Gulf of Mexico oysters and the estimated consumption of raw oysters together with the
monthly reported cases of V. vulnificus-associated septicaemia in that country. With further research,
this risk relationship will be applied in the V. vulnificus risk assessment and validated. Preliminary
results from this work were used in the risk characterization. However as this is a new approach to
developing a dose-response relationship it is currently being fine-tuned and therefore the doseresponse curve is not shown here.
The risk characterization linked the exposure assessment and the dose-response to predict
V. vulnificus illness rates. The predictions were compared to the observed illness rates. The model was
then used to evaluate targeted mitigation levels for risk reduction.
1. The FDA-VPRA provided a useful framework to model the risk of V. vulnificus septicaemia from
consumption of raw oysters.
2. The model predictions of V. vulnificus exposure were validated using independent data from a
survey of raw Gulf Coast oysters from the United States of America. Averaging water and air
temperature over a ten year period, as was done in the V. parahaemolyticus risk assessment, could
cause substantial deviations from observed levels in unusual climatic conditions such as the La
Nia that occurred in 1998. However, there was good agreement between observed and predicted
numbers of V. vulnificus using observed temperatures during this period as shown in Figure 6.7.
3. Preliminary results for the dose-response model indicated that closer agreement between
predicted and reported illness rates can be obtained by either eliminating data associated with
unusual climatic conditions or by using temperature, exposure and illness data individually for
each month of each year available (1995 to 2001) without averaging.
4. An aggregate population dose-response could be approximated using available data on the
differences in exposure to V. vulnificus from consumption of United States of America Gulf Coast
oysters and reported illness frequency during warm and cold months.
5. The approach for determining dose-response circumvents the lack of data on frequency of virulent
strains in raw oysters and uncertainty concerning the susceptible population by assuming that
these do not vary from month to month.
6. Preliminary results giving the predicted illness rate using the averaging of years approach
produced good agreement with observed illness rates except during the winter.
34
7. Preliminary evaluations of mitigations aimed at reducing V. vulnificus numbers in oysters to 3, 30

and 300 CFU/g indicate 60% to almost total reduction in predicted numbers of illness per year in
the United States of America. Although some further refining of the modelling is required it
appeared that this approach was appropriate for determining the potential effectiveness of specific
mitigations to reduce V. vulnificus illness associated with consumption of raw United States of
America Gulf Coast oysters.
Regional, seasonal & yearly variation

HARVES T
Water temperature
total Vv/g
POS T-HARVES T
Vv/g at harvest
Time to refrigeration
Air temperature
Vv/g at 1st refrigeration
Cooldown time
Vv/g at cooldown
S torage time
Vv/g (numbers) at consumption
Oysters per serving

Ingested dose
S usceptible population
Weight per oyster (g)

RIS K OF
ILLNES S
FIGURE 6.6: Schematic diagram of the V. vulnificus conceptual risk assessment model showing
integration of all the modules.

This risk assessment framework and in particular the dose-response relationship developed
using data for consumption of raw Gulf Coast oyster in the United States of America could be applied
to oysters and other molluscan shellfish species in other regions of the United States of America and
perhaps other countries. However, the use of data from this model as surrogate data for other regions
should be carefully considered, especially in conditions such as temperature and salinity that are
substantially different from those used to construct this model, and with shellfish species, culture
conditions or industry practices that are different than the submerged bottom culture that is typical of
the United States of America Gulf Coast. A modified temperature vs. V. vulnificus relationship for
high salinity (30-35 ppt) may be more appropriate for many parts of the world where shellfish
production is predominantly taking place in highly saline waters as the current model does not
incorporate salinity and overestimates V. vulnificus densities at high salinities. The model framework
35
is quite flexible and most model inputs could be easily adapted to fit specific situations if appropriate
data were available.
The dose-response approach used in this assessment was a curve-fitting of the data to a betapoisson model. While the beta-poisson model was selected any other empirical model that fits the data
could be used. Extrapolation of the beta-poisson parameters used in this analysis beyond the range of
normal consumption would be inappropriate.
The application of this model to predict the risk of V. vulnificus illnesses from seafoods other
than molluscan shellfish was limited as the ecology of this bacterium differs considerably as do
industry practices and consumer handling. However, the dose-response relationship could be useful in
determining the risk of other seafoods if V. vulnificus levels in these products were known at the point
consumption. The accuracy of these assessments would depend on the extent of matrix effects on the
dose-response.
The model does not account for variation of strain virulence. Regional or seasonal variation in
V. vulnificus virulence could alter the current dose-response and affect illness estimates.
This assessment was based on the distribution of at risk individuals52 in the population of the
United States of America and this parameter would need to be redefined on a country by country basis
depending on the size and characterization of the at risk population.
mean log MPN/g
observed retail study (June 1998- July 1999)

model prediction based on 1987-1997 average temperatures
model prediction based on Sept 1998 - Mar 1999 temperatures
-1
Winter
Spring
Summer
Fall
Season
FIGURE 6.7: Predicted and observed numbers of V. vulnificus per gram oyster on a seasonal basis.
Since the dose-response data was generated in part using monthly illness rates in the United
States of America, this data, in its current format, cannot be used to validate the model. However,
illness data from that country may be useful for validation of the risk characterization in a different
format (i.e. retrospective analysis of annual illness rates before and after specific mitigations). In 1997
the Interstate Seafood Sanitation Conference (ISSC) in the United States of America adopted a time52
Individuals with predisposing conditions which include insulin-dependant diabetes, liver disease (cirrhosis), gastric
acidity, cancer, hepatitis B & C, kidney disease, haemochromatosis, AIDS, being immunocompromised due to
treatment/surgery, asthma, rheumatoid arthritis, psoriatic arthritis, lupus, polymylagia rheumatica, giant cell arthritis and
being a transplant recipient.
36
temperature matrix for reducing the time to first refrigeration of oysters from 20 to 10h under certain
circumstances. The model could be used to analyse the effect on exposure and predicted illness and
these could be compared to illness rates before and after adoption of the time-temperature matrix.
In the course of this work the following data gaps were identified.
There was sufficient exposure data available for modelling the risk of V. vulnificus illness
from consumption of raw Gulf Coast oysters in the United States of America using the
proposed approach but this data especially numbers of V. vulnificus at harvest is also lacking
in most other countries.
There is a lack of reliable markers for virulence determination of V. vulnificus, thus requiring
the assumption that all strains are virulent.
The incidence of specific risk factors in the population consuming a seafood of interest and
exposure associated with this seafood are the primary data needed for applying this model to
other countries.
Validation of the model in a given region or country would require epidemiological data on
the incidence of primary septicaemia caused by V. vulnificus on a monthly basis.
6.1.7 Choleragenic Vibrio cholerae O1 and O139 in warm-water

shrimps for export
The justification for undertaking a risk assessment of this product-pathogen combination was
that shrimp is an important commodity in international trade and is occasionally suspected to be
involved in transmission of cholera, although there is little or no evidence that imported shrimp are
actually the vehicle of transmission. The total world shrimp production in 1999 was about four
million tons, of which 1.3 million tons were traded internationally with three quarters of this
originating from developing countries (FAO, 199953). Shrimp exports are negatively affected,
particularly when there are cases of cholera in shrimp producing countries.
A risk assessment of choleragenic V. cholerae O1 and O139 in shrimp for domestic use had
also been initiated. This was discontinued as shrimp consumed domestically does not appear to be an
important vehicle for transmission of cholera. Also, major difficulties and uncertainties exist in
defining handling and storage practices; possible routes of faecal cross-contamination and
consumption practices of domestic shrimp.
6.1.7.2 Scope
To assess the health risk of cholera associated with the consumption of imported warm-water
shrimp.
Toxigenic V. cholerae O1 and O139 are the causative agents of cholera, a water- and foodborne disease with epidemic and pandemic potential. Non-O1/non-O139 strains may also be
pathogenic but are not associated with epidemic disease. Non-O1/non-O139 strains are generally
nontoxigenic, usually cause a milder form of gastroenteritis than O1 and O139 strains, and are usually
associated with sporadic cases and small outbreaks rather than epidemics (Desmarchelier, 199754).
53
FAO (1999) World production of fish, crustaceans and molluscs reached 126.2 million tonnes in 1999, an increase of 7.2
% above the 1998 level. http://www.fao.org/fi/trends/worldprod99e.asp .
54
Desmarchelier, P.M. 1997. Pathogenic Vibrios. In A.D. Hocking, G. Arnold, I. Jenson, K. Newton and P. Sutherland, eds.
Foodborne Microorganisms of Public Health Significance 5th Edition, p 285 -312. North Sydney, Australian Institute of
Food Science and Technology Inc.
37
Outbreaks of cholera have been associated with consumption of seafood including oysters, crabs and
shrimp (Oliver and Kaper, 199755). The largest outbreak was a pandemic in South America in the
early 1990s when V. cholerae O1 caused more than 400,000 cases and 4,000 deaths, in Peru (Wolfe,
199256). Contaminated water used to prepare food, including the popular, lightly marinated fish
ceviche, was associated with the outbreak.
Cholera occurs in areas with inadequate sanitary conditions and infrastructure and is
associated with faecal contamination of water and foods. V. cholerae is widely distributed in coastal
and estuarine environments all over the world and there exists over 170 serotypes of V. cholerae.
According to WHO definitions57, only serotypes O1 and O139 are the causes of cholera.
Ability to produce cholera toxin (CT) is the determining virulence factor for causing cholera.
However, environmental strains of V. cholerae O1 have often been shown to be non-toxigenic.
Though some strains of non-O1/O139 V. cholerae may also cause gastroenteritis, the disease is of a
mild to moderate severity58. Choleragenic V. cholerae is susceptible to inactivation by cooking. Most
of the risk associated with choleragenic V. cholerae comes from the consumption of raw seafood or
from cross-contamination of the foods by food handlers or contaminated water.
Accordingly, this risk assessment considers only choleragenic V. cholerae O1 and O139.
V. cholerae O1 and O139 have both pathogenic and non-pathogenic forms based on the
presence of specific virulence genes, ctx (cholera toxin gene). Infection by choleragenic V. cholerae
O1 and O139 is characterized by an acute gastroenteritis. Therefore, the end-point of the doseresponse curve was defined as gastroenteritis.
Human volunteer studies were available for the construction of the dose-response curves for
V. cholerae O1. Reasonable Beta-Poisson dose-response parameters were obtained from data sets;
however, the human volunteer studies characterize dose-response relationships for pathogens
administered with a pH-neutralizing buffer rather than for pathogens administered with a food matrix.
In normochlorohydric adult volunteers, doses of up to 10 11 choleragenic V. cholerae given without
buffer or food did not reliably cause illness, whereas doses of 10 4 to 10 8 organisms given with
NaHCO3 (sodium bicarbonate) resulted in diarrhoea in 90% of individuals. Dose-response curves
(Figure 6.8) show that a high dose of V. cholerae O1 (10 6) was normally needed to cause illness
when V. cholerae are consumed in food. In populations not exposed to choleragenic V. cholerae all
age groups are equally susceptible. Immunity seems to be serotype specific.
This risk assessment includes aquacultured and wild-caught warm-water shrimp.
Choleragenic V. cholerae O1 and O139 generally occur in waters with salinity's between 0.2 to 20
ppt. Therefore, water and shrimp from offshore waters have not been found to contain choleragenic
V. cholerae. Thus, it is assumed that any presence of choleragenic V. cholerae in offshore wild caught
shrimp is caused by post-harvest cross-contamination. Though the presence of choleragenic
V. cholerae in aquaculture environments is very rare, the model assumes that such choleragenic
V. cholerae strains could be present in shrimp at levels similar to those found in coastal waters of
cholera endemic countries.
The model developed was based on shrimp handling, processing and storage practices in units
approved for export of shrimp (Figure 6.9). Such approval is based on sanitary requirements as
described in Good Manufacturing Practices (GMP) and Good Hygienic Practices (GHP). Shrimp
55
56
Wolfe, M. 1992. The effects of cholera on the importation of foods: Peru- a case study. PHLS Microbiology Digest, 9: 4244.
57
WHO Fact Sheet N107
58
Morris, J.G. Jn. 1990. Non-O group 1 Vibrio cholerae: a look at the epidemiology of an occasional pathogen.
Epidemiological Review, 12: 179-191.
38
intended for export are generally iced immediately after harvest and transported in ice to certified
processing units that meet GHP/GMP requirements. However, a worst-case scenario of shrimp being
processed in non-approved units was also considered.
The major factors influencing the numbers of choleragenic V. cholerae in shrimp are time and
temperature during handling, processing and storage. In the absence of available data it was necessary
to make assumptions on distributions of time and temperature under such conditions. Adequate data
were available on the effect of washing, freezing and cooking on the numbers of choleragenic
V. cholerae in shrimp. In particular, the duration of frozen storage before consumption will cause a
significant reduction in numbers of choleragenic V. cholerae. Limited information was available on
the levels of faecal cross-contamination during handling and multiplication of choleragenic
V. cholerae in raw shrimp. The model takes into account the pronounced reduction in the levels of
choleragenic V. cholerae that would occur during cooking of shrimp either before export or before
consumption. It also assesses the risk if shrimp were consumed raw or inadequately cooked in the
importing country.
1.00
probability of illness
0.80
Classical with Food Matrix
(Cash)
El Tor (Levine 1988)
0.60
Classical with antacid (Cash)

misc El Tor strains tested
0.40
fit to Classical w food matrix

fit to El Tor with antacid
0.20
fit to Classical Inaba

Classical Ogawa (Cash)
0.00
1.E+00
1.E+02
1.E+04
1.E+06
1.E+08
1.E+10
1.E+12
cholera dose
FIGURE 6.8: Beta poisson does-response curve for Vibrio cholerae

The risk characterization was undertaken by combining the dose-response model with the
estimated exposure to choleragenic V. cholerae via shrimp. Based on the available data, including
additional information that was identified by the expert consultation, but not yet considered in the risk
assessment, it will be feasible to progress with a semi-quantitative risk assessment. Further,
epidemiological data on cholera cases reported to the WHO from major countries importing warmwater shrimp is available and this, together with data on shrimp imports and consumption, will be
collected to validate the model.
1. Following an extensive literature review it was noted that while V. cholerae is widely distributed
in the environment, only strains producing cholera toxin and belonging to serotypes O1 and O139
are causative agents of cholera.
39
2. Contamination of shrimp, either wild-caught offshore or aquacultured, with choleragenic

V. cholerae could happen during handling and processing, but there is very little opportunity for
the multiplication of V. cholerae in shrimp processed in units meeting GMP/GHP requirements.
3. Major log-reductions in numbers of choleragenic organisms occur during washing, freezing and
cooking.
4. Dose-response curves (Figure 6.8 ) show that a high dose of choleragenic V. cholerae O1 (10 6) is
normally needed to cause illness when choleragenic V. cholerae O1 are consumed in food.
5. The qualitative (descriptive) risk assessment showed that there was not a public health problem
associated with the consumption of imported warm-water shrimp.
Shrimp harvested from

coastal areas/wild
caught or
acquaculture ponds
V. cholerae / g
V. cholerae / cm on
fingers of handlers
V. cholerae / g in ice
V. cholerae / ml in
water
Shrimp washed, iced

and transported to
processing
establishments
V. cholerae / ml in water
Shrimp washed,
peeled, graded,
packed and frozen
V. cholerae / g
Time and temperature of

frozen storage
Shrimp in frozen condition

in supermarkets
V. cholerae / g
Time and temperature of
frozen storage
Shrimp thawed, cooked

and consumed
No. of V. cholerae
ingested
FIGURE 6.9: Conceptual model for risk assessment of choleragenic V. cholerae in warm water
shrimp for export.
40
6.1.7.8 Limitations and caveats

There was limited or negative data available on the level of choleragenic V. cholerae O1 and
O139 in shrimp at harvest. Estimations were based on reported levels found in waters.
Only one 20-year old reference was available on the lack of multiplication of choleragenic
V. cholerae in raw shrimp.
There was no data on the level of choleragenic V. cholerae that may be transmitted by shrimp
handlers, e.g. on fingers. Therefore, an assumption had to be made on transmission of V. cholerae by
faecal cross-contamination.
The dose-response data for choleragenic V. cholerae O1 consumed with food were available
for the classical biotype but not for the El Tor biotype. The El Tor biotype is the more common form
and dose-response information on this form when administered with food rather than acid neutralizing
vehicles is desirable.
The following data gaps were identified in the course of the work.
Data on the levels of choleragenic V. cholerae O1 and O139 in natural waters and aquaculture
environments.
Data on the multiplication of choleragenic V. cholerae in cooked and raw shrimp.
Data on levels of faecal cross contamination during handling of shrimp.
Data to clarify the dose-response when the El Tor form is ingested in food.
6.2 Review of the Risk Assessments

The expert consultation undertook a technical review of the draft risk assessment document
entitled A Draft Risk Assessment of Vibrio spp. in Seafood. The expert consultation evaluated the
risk characterization, as well as the underlying data, assumptions, and associated uncertainty and
variability. The expert consultation recognized the extensive work undertaken by the drafting group,
provided additional references to meet some of the specific data needs of the risk assessment and
made recommendations on how to improve the document.
6.2.1 Introduction: Vibrio spp. in seafood

The expert consultation noted that it was desirable to state that the pathogenicity of
V. parahaemolyticus is associated with TDH and/or TRH production. With regard to V. cholerae, it is
necessary to state clearly that epidemic cholera is only associated with cholera toxin-producing strains
of serogroups O1 and O139. It was recognized that not all professionals may be aware that virulence
of V. parahaemolyticus, and O1 and O139 V. cholerae is associated with certain toxin-encoding
genes, and that diagnostic tests can be used to separate these strains from others.
The expert consultation expressed concern that testing of seafood for Vibrio spp. was
sometimes based on inappropriate markers (e.g. genus, species and/or non-consideration of
pathogenic factors) that do not reflect the potential to cause human illness. It was agreed that the risk
assessment should include a section outlining the Vibrio spp., types and virulence factors that may be
included in the examination of different types of seafood in order to protect public health.
41
6.2.2 Vibrio parahaemolyticus in raw oysters consumed in Japan,

New Zealand, Canada, Australia and the United States of
America oysters
The expert consultation identified that the risk assessment should include further
consideration of the oyster industry practices in different parts of the world as these could have a
significant effect on the appropriateness of the present model. In particular, this applied to the
uncommon use of refrigerated transport and storage in many countries. The caveats that applied to the
assessment due to these differences should be specified. It was determined that modelling should also
consider salinity as a parameter, since in some areas of the world, salinity remains high throughout the
year and exerts an effect in addition to that of seawater temperature. This might be addressed by
having separate models for areas of relatively constant high salinity; relatively constant low salinity;
and varying salinity. It was also necessary to consider that different oyster species might behave
differently with regard to both the concentration of V. parahaemolyticus in the harvest area and to the
growth of the organism during periods of temperature abuse or cool-down. Several of these
considerations could have led to the model over-predicting the incidence of illness due to
V. parahaemolyticus in oysters in Australia and New Zealand.
It was highlighted that the model did not currently incorporate consideration of TRHproducing strains of V. parahaemolyticus, nor did it presently encompass possible variability in the
prevalence of TDH-producing strains seen in other countries. The model could be amended in the
future to address these aspects.
The risk assessment should include fuller consideration of uncertainty and variability and it
would be useful to include an outline of the use of the @Risk model. It would be necessary to specify
as fully as possible the data needed, as well as any key caveats, if the model were to be applied or
modified to be used in other regions and for other species.
6.2.3 Vibrio parahaemolyticus in bloody clams

The expert consultation congratulated the drafting group and the associated researchers for
undertaking a valuable targeted risk assessment in a very short period of 5 months, going from
literature review and capture of novel data to modelling and drafting of the assessment. The process
itself could form an example for future targeted assessments.
The expert consultation recommended that the drafting group revise the assessment itself to
ensure clarity with regard to the data and methods used (both microbiological and modelling) and to
ensure that the figures and other outputs were sufficiently explained in the text. There was the
potential to include more about the uncertainty and variability in the model and to look in more detail
at the modelling aspects of the mitigation process (boiling). These recommendations did not detract
from the quality of the work that has already been done.
6.2.4 Vibrio parahaemolyticus in finfish

An exposure assessment had been prepared previously and presented to the first expert
consultation in Geneva in 200159. FAO and WHO, in conjunction with the drafting group,
subsequently decided that there were insufficient data to take the assessment forward at the present
time and therefore no further work had been undertaken. The expert consultation recognized the
useful content included in the exposure assessment and determined that it should be included in the
final report on the Vibrio risk assessments and could then form the basis for possible future work and
as support to FAO/WHO member countries and Codex.
59
42
6.2.5 Vibrio vulnificus in raw oysters

The expert consultation noted the success in extending the use of the V. parahaemolyticus risk
assessment and model to another species. It was discussed that the models are currently based on
United States of America Gulf Coast data sets, and that the illness data used for validation are from
the same country. The development of an additional salinity-temperature model will assist in
extending this tool to other environments where high salinity may be a factor in limiting exposure.
The stipulations noted for the V. parahaemolyticus assessment also apply to this assessment and
appropriate data needs and caveats for application to other regions also need to be emphasized. The
assessment should also include further discussion as to how the tool can be used in regions where
shellfish -associated V. vulnificus infection may be of importance.
6.2.6 Choleragenic Vibrio cholerae O1 and O139 in warm-water

shrimp for export
According to WHO definitions60, only serotypes O1 and O139 are the causes of cholera and
the ability to produce cholera toxin (CT) is the determining virulence factor. However, environmental
strains of V. cholerae O1 have often been shown to be non-toxigenic, therefore, according to the
expert consultation, seafood products should only be analysed for cholera toxin - producing
V. cholerae O1 and O139.
The adaptation of the V. parahaemolyticus model in oysters (based on temperatureV. parahaemolyticus numbers predictor) to a V. cholerae O1/O139 choleragenic model in warm-water
shrimp was recognized as difficult because of the absence of a good predictor for V. cholerae numbers
in shrimp, and it would therefore require a large amount of work to develop a completely new model.
The export of warm-water shrimp constitutes an important aspect of world trade. It was
generally agreed that the available qualitative (descriptive) risk assessment showed that there was not
a public health problem associated with the consumption of imported warm-water shrimp; however a
semi-quantitative risk assessment should be undertaken with the available data in order to assist risk
managers to better understand this.
It was decided not to proceed with a risk assessment of shrimp consumed by domestic
markets in tropical countries. Since shrimp are consumed cooked, any illness will be the result of
cross-contamination and this is not specific to any single food commodity.
6.3 Utility and applicability

The expert consultation emphasized the need for the risk assessments to be distributed as
widely as possible and in forms that are appropriate to the target groups. As well as the planned
Executive Summary and full Technical Report, an Interpretative Summary should be produced that
explains the use and limitations of these risk-based tools without great emphasis on details of the
models. FAO and WHO should consider the publication of entire consultation reports in respected,
widely read journals. Some issues that need to be considered here are FAO and WHO policies on
copyright, the need for peer review of already heavily reviewed documents and the timing of
publication. In addition, the drafting group should be encouraged to submit relevant aspects of the
work for publication in peer-reviewed journals to reach a greater audience. It will be important to
consider publication of elements prior to publication of the full report in order to contribute to general
development of microbiological risk assessment for foods.
It will be necessary for presentations to be made to relevant stakeholders, including Codex, in
order to ensure that the relevant aspects were fully emphasized. Provision of appropriate PowerPoint
presentations by FAO and WHO would assist in this. It was also perceived that the results of the work
would be of value to the seafood industry and thus simplified summaries of the assessments should be
submitted to trade periodicals. The benefits of these risk assessments will be realized through the use
of trainers who are skilled in communicating with diverse types of audiences. FAO and WHO should
60
WHO Fact Sheet N107
43
make the software models available together with guidance on their use. They should also provide
technical assistance to developing countries wishing to extend the assessments to local needs.
These risk assessments could be used to support relevant risk management decisions. The
output from the assessments should also be used to formulate and target research needs, e.g. to fill
identified data gaps.
6.4
Response to the specific questions posed by the Codex

Committee on Fish and Fishery Products
A number of questions were posed to the expert consultation in relation to management

strategies for food-borne illness due to V. parahaemolyticus and V. vulnificus. These were posed to
the members of the consultation in their roles as experts in the microbiology of vibrios and/or seafood
technology. The four questions posed by the CCFFP and the response from the expert consultation are
outlined below.
Question 1: Whether the following pre-harvest control measures (testing/monitoring the
following parameters and consequential closure of the harvesting area) are effective in the control of
Vibrio parahaemolyticus and Vibrio vulnificus in bivalve molluscs:
Testing of bivalve mollusc meat for Vibrio parahaemolyticus and Vibrio vulnificus
Temperature monitoring of the growing water
Water testing for Vibrio parahaemolyticus and Vibrio vulnificus
Salinity monitoring
Response from the consultation: The concentrations of V. parahaemolyticus and
V. vulnificus in shellfish may be measured directly or predicted by monitoring temperature and
salinity. There will not necessarily be a direct relationship between these surrogate variables and the
measured concentrations of pathogenic vibrios for a particular area as there is uncertainty and
variability in the current models. The predictive abilities of the models would be improved by
incorporating local data and considering additional factors such as hydrodynamic effects and sunlight.
The effectiveness of these measures in controlling illness would depend on the instigation of an
appropriate mitigation (or multiple mitigations) and this is not confined to closure of a harvesting
area.
The current models do not include modules relating to the concentration of the two pathogens
in seawater and thus the utility of measuring this cannot be estimated. If the appropriate data were
gathered then the models could be extended accordingly.
Question 2 Are the following post-harvest treatment technologies, alone or in combination,
effective in the reduction or elimination of V. parahaemolyticus and V. vulnificus in bivalve molluscs:
hydrostatic pressure
rapid cooling
irradiation
mild heat treatment (pasteurization)
freezing and thawing
depuration
Response from the consultation: These may all have the effect of reducing the numbers of
pathogenic vibrios but the effectiveness will vary according to the conditions of use, and there may be
a need to balance between obtaining the maximum possible reduction in bacterial content and
retaining consumer-acceptance of either the product or the process. Reports on the effectiveness of
depuration vary greatly and this may again depend on the conditions of use - some reports indicate
that proliferation of vibrios may occur during this process. The general opinion of the expert
consultation is shown on a qualitative/semi-quantitative basis in table 6.7.
The current models could be adapted to enable estimates to be obtained of the effectiveness of
the mitigations in reducing illness. With regard to the mitigation of the closure of harvesting areas,
44
estimates could also be obtained of the proportion of harvest lost by application of a particular
scenario.
Some of the listed mitigations are also used in combination, e.g. hydrostatic pressure and
freezing; depuration and hydrostatic pressure or pasteurization.
TABLE 6.7: The comparative effectiveness of a number of mitigation strategies in reducing Vibrio
spp..
Mitigation
Hydrostatic pressure
Rapid cooling
Irradiation
Pasteurization
Freezing and thawing
Depuration
Relay at high salinity for 2 weeks
(for V. vulnificus)
Commercial heat-treatment
+
++
+++
Comparative effectiveness in reducing

Vibrio spp.
+++
+/++
+++
+++
++
+/++
+++
no effect
some reduction
moderate reduction
significant reduction
References and further reading relating to inactivation strategies

Calik, H., Morrissey, M.T., Reno, P.W. & An H. 2002. Effect of high pressure processing on Vibrio
parahaemolyticus strains in oysters. Journal of Food Science, 67: 1506-1510.
Cook, D. W., and A. D. Ruple. 1992. Cold storage and mild heat treatment as processing aids to
reduce the numbers of Vibrio vulnificus in raw oysters. Journal of Food Protection, 55:985989.
Cook, D. W., and A. D. Ruple. 1989. Indicator bacteria and Vibrionaceae multiplication in postharvest shellstock oysters. Journal of Food Protection, 52:343-349.
Cook, D. W. 1999. Effect of heat and freezing treatment on Vibrio parahaemolyticus O3:K6.
Unpublished data.
Eyles, M. J., and G. R. Davey. 1984. Microbiology of commercial depuration of the Sydney rock
oyster, Crassostrea commercialis. Journal of Food Protection, 47:703-706.
Gooch, J. A., A. DePaola, C. A. Kaysner, and D. L. Marshall. 1999. Postharvest growth and survival
of Vibrio parahaemolyticus in oysters stored at 26 C and 3 C. Abstracts of the 99th General
Meeting of the American Society for Microbiology, Abstract # P52.:521.
Greenberg, E. P., and M. Duboise. 1981. Persistence of Vibrio parahaemolyticus and Vibrio harveyi
in hardshell clams. Abstracts of the 81st General Meeting of the American Society for
Microbiology, Abstract No. Q93.:216.
Johnson, H. C., and J. Liston 1973. Sensitivity of Vibrio parahaemolyticus to cold in oysters, fish
fillets and crabmeat. Journal of Food Science, 38:437-441.
Johnson, W. G., Jr., A. C. Salinger, and W. C. King. 1973. Survival of Vibrio parahaemolyticus in
oyster shellstock at two different storage temperatures. Applied Microbiology, 26:122-123.
Motes, M.L. and DePaola, A. 1996. Offshore suspension relaying to reduce levels of Vibrio vulnificus
in oysters (Crassostrea virginica). Applied and Environmental Microbiology, 62, 3875-3877.
45
Richards, G. P. 1988. Microbial purification of shellfish: A review of depuration and relaying. J. Food
Protect. 51:218-251.
Son, N. T., and G. H. Fleet. 1980. Behaviour of pathogenic bacteria in the oyster, Crassostrea
commercialis, during depuration, re-laying, and storage. Applied and Environmental
Microbiology, 40:994-1002.
Question 3
For Vibrio parahaemolyticus - Are food-borne illnesses caused by the heat
resistant toxin produced by the pathogen or by the pathogen itself?
Response from the consultation: The illness is caused by the toxin but only if this is
produced in the intestine following colonization by a strain producing TDH, TRH or both toxins.
Question 4: What is the availability of methods of analysis for Vibrio parahaemolyticus
toxin gene (tdh)?
Response from the consultation: Both tdh and trh genes can be detected using PCR with
relevant primers and by membrane filtration-hybridization methods with non-isotopic oligonucleotide
or PCR-generated probes. For quantification, PCR methods can be applied in an MPN format whereas
membrane filter-hybridization can be used for direct colony enumeration. PCR and colony
hybridization procedures are also available for the thermolabile haemolysin gene (tlh) for determining
V. parahaemolyticus species. As with conventional methods, there is scope for standardization and/or
the determination of the relative performance of current methods.
6.5 Response to the needs of the Codex Committee on Food

Hygiene
A CCFH drafting group has prepared a document titled Discussion Paper on Risk
Management Strategies for Vibrio spp. in Seafood. In that paper, some risk assessment needs and
questions for risk assessors were identified, which include an evaluation of the impact of several
potential interventions on the risk of V. parahaemolyticus infection. The current risk assessment of
Vibrio spp. in seafood is addressing many of these potential interventions through the models that
have been developed including the influence of temperature on growth and the impact of various
target reduction levels of Vibrio spp. in oysters on the risk of illness. The effects of various mitigation
strategies are also described in section 6.4.
6.6 Conclusions and recommendations

The drafting group has progressed towards improving the risk assessments since the last
expert consultation61. The majority of the previous recommendations have been addressed in the
current draft report.
The vibrio risk assessments should include clear advice as to the species, serogroups,
serotypes and genotypes that should be considered as being of public health significance with respect
to the trade and consumption of seafood. Interpretative summaries of the risk assessments should be
produced in order to maximise the understanding of the work by other professionals. Where
appropriate, these summaries should have limited descriptions of the mathematical modelling in order
to reach a wider audience.
It was recommended that the risk assessment on "Vibrio cholerae in shrimp" be developed
further on semi-quantitative basis only and the opinions of risk managers should be sought before any
work is undertaken towards a fully quantitative risk assessment.
61
WHO 2001. Report of the Joint FAO/WHO Expert Consultation on Risk Assessment of Microbiological Hazards in
46
The exposure assessment on "Vibrio parahaemolyticus in finfish" should be included in the

final report on the vibrio risk assessments as this contains information that may be useful to a number
of countries.
The procedures used to undertake the risk assessment on bloody clams should be recognized
as a way to expedite the development of pathogen-commodity risk model. The draft of the risk
assessment itself should be revised to ensure greater transparency of the modelling approaches.
The risk assessment on "Vibrio parahaemolyticus in oysters" should include further
consideration of the oyster industry around the world (practices, species effects, harvest water salinity,
etc). Where the model cannot be modified appropriately, the potential constraints and/or the additional
data needs should be clearly identified. It should identify the potential shortcomings of the assumption
which precluded the consideration of TRH-producing strains as pathogenic and also the possible
consequences of the high prevalence of TDH-producing strains in some parts of the world. Further
consideration should be given to uncertainty and variability, and it should also include an description
of the Excel-based model in order to assist understanding its potential applications and limitations.
The risk assessment on "Vibrio vulnificus in oysters" should include consideration of the
potential constraints and/or the additional data needs to extend it to other geographical areas outside
of the United States of America. It should identify more clearly those areas of the world and the
seafood products that were currently known to be associated with food-borne V. vulnificus infection.
In order to undertake further risk assessments it will be necessary to obtain additional
information on the proportion of strains of each Vibrio spp that possess pathogenic traits. Virulence
factors of V. parahaemolyticus (TDH and TRH) and V. cholerae O1 and O139 (CT) should be
identified where appropriate so as to specify the strains that are relevant to human illness. The expert
consultation believed that additional information on dose-response relationships with respect to food
contaminated with Vibrio spp. could be obtained by investigation of outbreaks.
7 Conclusions
The assessments presented at this expert consultation were in varying degrees of completion.
The risk assessments for Campylobacter spp. in broiler chickens, V. vulnificus and
V. parahaemolyticus in oysters, are most suited at the present time for use by risk managers to help
them make informed risk management decisions. The models use a modular approach which lends
flexibility for their use in assessing production systems not specifically covered in the current models.
The utilization of the models was demonstrated by a number of examples showing how certain
mitigation strategies may lead to a reduction in the number of cases of disease. The models are still
evolving and once fully completed will become a useful tool for risk managers.
Since the last consultation, the hazard characterization and exposure assessment were merged
to produce a preliminary risk characterization. The risk characterization can provide risk managers
with the ability to gain insight into the relative effectiveness of various mitigation strategies. Future
activities will focus on ensuring the full potential of the risk assessment is achieved. In addition, a full
exploration of the model will be undertaken to fully understand the scope and limitations
8 Recommendations
The expert consultation recommended that FAO and WHO should:

ensure that for future risk assessment activities, a clear-cut statement of purpose be provided by
the risk managers at the start of risk assessment activities, followed by an active and continuous
interaction between risk managers and risk assessors. It is important that risk managers are
involved in the design of the risk assessment and assist in making explicit the scope of the
assessment. This will ensure that the outcomes will be of maximum use to all parties.
highlight the need to collect quantitative data to all steps of the risk assessments, so as to provide
appropriate risk management conclusions.
47
provide assistance to developing countries to gather quantitative information for risk assessment.
Appropriate resources should be provided to developing countries to train and undertake risk
assessments and those wishing to extend the risk assessments to local needs.
encourage microbiologists to develop and employ techniques to differentiate pathogenic and nonpathogenic strains.
encourage governments that have set or intend to set targets for food-borne diseases addressed in
this report to use the models presented to inform their decisions in setting appropriate food safety
objectives and intervention strategies.
commission a document to assist a wider readership in understanding this report. The
recommended document would describe for non-specialists appropriate information on;
i) shellfish and poultry e.g. production methods, processing, economics and legislation.
ii) Vibrio spp. and Campylobacter spp. (surveillance, epidemiology and microbiology).
iii) risk assessment (its approach, components and how it can answer risk managers
questions).
facilitate hands-on demonstrations for identifying and collecting relevant data for risk assessment
for specific commodity/pathogen combinations including risk characterization using various
modelling software.
develop new ways of communicating the concepts of risk assessments to users.
promote utilization of the valuable framework models that have now been established to
undertake further risk assessments. It would be relatively easy to use modules from these risk
assessments to develop a risk assessment of Salmonella in shrimps. The experts consider this
would be a valuable risk assessment for future consideration.
consider, for future risk assessment activities, the involvement of scientific experts to serve as a
standing advisory resource group to the risk assessment drafting group, for the duration of a
specific work assignment. This would be a significant complementary activity to the more formal
in-person experts meetings and would certainly take greater advantage of the intellectual
resources of the expert community through their knowledge of and access to relevant data,
references and network of colleagues.
encourage the submission of appropriate aspects of the work to peer-reviewed technical
publications and the seafood and poultry trade press.
make the software models available in a format usable by other professionals and provide
appropriate guidance on their use.
48
Annex 1: List of participants

INVITED EXPERTS
Nourredine Bouchriti, Department d'Hygine et d'Industrie des Denres Alimentaires d'Origine Animale,
Institut Agronomique et Vtrinaire Hassan II, B.P. 6202, Instituts, 10101 Rabat, Morocco
John Cowden, Scottish Centre for Infection and Environmental Health, Clifton House, Clifton Place, Glasgow
G3 7LN, United Kingdom
Heriberto Fernndez, Instituto de Microbiologa Clnica, Universidad Austral de Chile, PO box 567 Valdivia, Chile
Jean-Michel Fournier, Unit du Cholra et des Vibrions, Centre National de Rfrence des Vibrions et du
Cholra, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15 France
Ron Lee, CEFAS Weymouth Laboratory, Barrack Road, The Nothe, Weymouth Dorset DT4 8UB, United
Kingdom
Carlos Lima dos Santos, Rua E. Souza Gomes 510/Cob, 22620-320 Rio de Janeiro, Brazil
Dorothy-Jean McCoubrey, Ministry of Agriculture and Forestry, P O Box 1254, Auckland, New Zealand
Geoffrey Mead, Private Consultant, 17 Harbutts, Bathampton, Bath, BA2 6TA Somerset, United Kingdom
Marianne Miliotis, FDA/CFSAN HFS-006, 5100 Paint Branch Parkway, College Park, MD 20740-3835,
United States of America
Diane G. Newell, Veterinary Laboratories Agency (Weybridge), New Haw, Addlestone, Surrey KT 15 3NB,
United Kingdom
Mitsuaki Nishibuchi, Center for Southeast Asian Studies, Kyoto University, 46 Shimoadachi-cho, Yoshida,
Sakyo-ku, Kyoto 606-8501, Japan
Serv Notermans, TNO Nutrition and Food Research Institute, Zeist, The Netherlands
Pensri Rodma, Department of Medical Science, Ministry of Public Health, 88/7 Tivanonth Rd, Soi
Bumrajnaradul, 11000 Bangkok, Thailand
Sasitorn Kanarat, Veterinary Public Health Laboratory, Division of Veterinary Public Health, Department
Livestock Development, Phayathai Rd. Phayathai, 10400 Bangkok, Thailand
Mark Tamplin, Microbial Food Safety Research Units, North Atlantic Area Eastern Regional, Research Centre
ARS, USDA, 600 East Mermaid Lane Wyndmoor, Pennsylvania 19038-8598, United States of America
Paul Vanderlinde, Senior Microbiologist, Food Science Australia, PO Box 3312, Tingalpa DC, Queensland
4173, Australia
DRAFTING GROUP EXPERTS: CAMPYLOBACTER SPP. IN BROILER CHICKENS
Bjarke Bak Christensen, Danish Veterinary and Food Administration, Institute of Food Safety and
Toxicology, Division of Microbiological Safety, 19, Morkhoj Bygade, 1860 Soborg, Denmark
Aamir Fazil, Population and Public Health Branch, Health Canada, 110 Stone Road West, Guelph, Ontario
N1G 3W4, Canada
Emma Hartnett, Department of Risk Research, VLA (Weybridge), Surrey, KT15 3NB United Kingdom
Greg Paoli, Decisionalysis Risk Consultants, Inc., 1831 Yale Avenue, Ottawa, Ontario Canada K1H 6S3
Maarten Nauta, Microbiological Laboratory for Health Protection (MGB), National Institute for Public Health
and the Environment (RIVM), P.O. box 1, 3720 BA Bilthoven, The Netherlands
Anna Lammerding, Population and Public Health Branch, Health Canada, 110 Stone Road West, Guelph,
Ontario N1G3W4, Canada
49
DRAFTING GROUP EXPERTS: VIBRIO SPP. IN SEAFOOD

Anders Dalsgaard, Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University,
Stigbjlen 4, DK-1870 Frederiksberg C, Denmark
Angelo DePaola, Office of Seafood, CFSAN, USFDA, Dauphin Island, AL, United States of America
Thomas McMeekin, Centre for Food Safety and Quality, School of Agricultural Science/Tasmanian Institute of
Agricultural Research, University of Tasmania, GPO Box 252-54, Hobart TAS 7001 Australia
Ken Osaka, National Institute of Infectious Diseases, Ministry of Health, Labour and Welfare 1-23-1 Toyama,
Shinjuku-ku, Tokyo 162-8640, Japan
John Sumner, M&S Food Consultants Pty. Ltd., Deviot Road, Deviot 7275, Australia
Mark Walderhaug, HFS-517, Center for Food Safety and Applied Nutrition, U.S. FDA, 5100 Paint Branch
Parkway, College Park, Maryland 20740-3835, United States of America
I. Karunasagar, Department of Fishery Microbiology College of Fisheries, University of Agricultural Sciences,
PB No. 527, Mangalore 575-002, Karnataka, India
JOINT FAO/WHO SECRETARIAT
Maria de Lourdes Costarrica, Senior Officer, Food Quality Liaison Group, Food Quality and Standards
Service, Food and Nutrition Division, Food and Agriculture Organization of the United Nations, Rome, Italy
Sarah Cahill, Food Quality Liaison Group, Food Quality and Standards Service, Food and Nutrition Division,
Food and Agriculture Organization of the United Nations, Rome, Italy
Lahsen Ababouch, Chief, Fish Utilization and Marketing Service, Fishery Industries Division, Fisheries
Department, Food and Agriculture Organization of the United Nations, Rome, Italy
Peter Karim BenEmbarek, Food Safety Programme, Department of Protection of the Human Environment,
World Health Organization, 20 Avenue Appia, CH-1211 Geneva 27, Switzerland
Hajime Toyofuku, Food Safety Programme, Department of Protection of the Human Environment, World
Health Organization, 20 Avenue Appia, CH-1211 Geneva 27, Switzerland
Jeronimas Maskeliunas, Food Standards Officer, Joint FAO/WHO Food Standards Programme, Food and
Nutrition Division, Food and Agriculture Organization of the United Nations (FAO), Viale delle Terme de
Caracalla, 00100 Rome, Italy
Karen Hulebak, Acting Deputy Administrator for Public Health and Science, United States Department of
Agriculture, Food Safety and Inspection Service, Rm 341E, 1400 Independence Ave, SW, Washington, DC
20250-3700, United States of America
50
Annex 2: List of working documents

Two working papers were prepared for, and presented during the expert consultation. These
served as the basis for the discussions, which led to the development of the report and the
recommendations. These documents were prepared for FAO and WHO by a number of expert drafting
groups. The full text of these documents will be made available on the FAO and WHO webpages;
http://www.fao.org/es/esn/food/risk_mra_en.stm and http://www.who.int/fsf/Micro/index.htm.
Paper no.
Title
Authors
(in alphabetical order)
MRA 02/01
A draft risk assessment

of Campylobacter spp.
in broiler chickens
Steve Anderson, Food and Drug Administration, United States of America

Bjarke Bak Christensen, Veterinary and Food Administration, Denmark
Aamir Fazil, Health Canada, Canada
Emma Hartnett, Veterinary Laboratories Agency, United Kingdom
Anna Lammerding, Health Canada, Canada
Maarten Nauta, Rijksinstituut voor Volkgesondheid en Milieu (RIVM), The
Netherlands
Greg Paoli, Decisionalysis Risk Consultants, Canada
Hanne Rosenquist, Veterinary and Food Administration, Denmark
MRA 02/02
A draft risk assessment

of Vibrio spp. in
seafoods
John Bowers, Food and Drug Administration, United States

Anders Dalsgaard, Royal Veterinary and Agricultural University, Denmark
Angelo DePaola, Food and Drug Administration, United States
I. Karunasagar, University of Agriculture Sciences, India
Ken Osaka, National Institute of Infectious Diseases, Japan
Thomas McMeekin, University of Tasmania, Australia
John Sumner, M&S Food Consultants Pty. Ltd., Australia
Mark Walderhaug, Food and Drug Administration, United States
51

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Risk assessment

and Vibrio spp. in seafood

Report of a Joint FAO/WHO Expert

OBJECTIVES OF THE CONSULTATION .............................................................................................2

SUMMARY OF THE GENERAL DISCUSSIONS...................................................................................3

RISK ASSESSMENT OF THERMOPHILIC CAMPYLOBACTER SPP. IN BROILER CHICKENS4

5.1.1 Summary of the Risk Assessment.......................................................................................................4

6.3 UTILITY AND APPLICABILITY ....................................................................................................................43

ANNEX 1: LIST OF PARTICIPANTS.............................................................................................................49

Codex Alimentarius Commission

3 Objectives of the consultation

the assumptions made in the risk assessment;

the associated uncertainty and variability;

the potential application of the work.

To respond to the risk management needs of Codex.

4 Summary of the general discussions

Expert members of the drafting group

Members of the secretariat

Vibrio spp. in seafood

Expert members of the drafting group

Members of the secretariat

5 Risk Assessment of thermophilic Campylobacter spp. in

6. Defeathering (an increase the magnitude of cross-contamination)

Between flock refers to different groups of hens (i.e. different flocks).

Skewness is a statistical term that refers to the lobsideness of a distribution.

5.1.5 Key findings

5. Overall campylobacter concentration on chicken carcasses decreased through processing, with

5.1.6 Risk assessment and developing countries

5.1.7 Limitations and caveats

5.1.8 Gaps in the data

Data on strain variability regarding virulence/pathogenicity and survival during

5.2 Review of the Risk Assessment

5.2.2 Hazard identification

5.2.3 Exposure assessment:

5.2.4 Hazard characterization

5.2.5 Risk characterization

5.2.6 Risk assessment and developing countries

5.2.7 Data deficiencies

5.3 Utility and Applicability

5.4 Response to the specific risk management questions posed by

5.5 Conclusions and Recommendations

6 Risk assessment of Vibrio spp. in seafood

6.1.3 Vibrio parahaemolyticus in raw oysters consumed in Japan,

log dose of V.parahaemolyticus

FIGURE 6.1: Beta-Poisson dose-response curve for V. parahaemolyticus (endpoint modelled is

6.1.3.5 Exposure assessment

R egional, seasonal & yearly variation

W ater tem perature

FIGURE 6.2:Harvest module for exposure assessment of V. parahaemolyticus in oysters.

V p/g at con su m ption

FIGURE 6.3: Post-harvest module for exposure assessment of V. parahaemolyticus in oysters.

FIGURE 6.4: Consumption module for exposure assessment of V. parahaemolyticus in oysters

6.1.3.6 Risk characterization

Japanese water temperature is available at http://www.hiroins-net.ne.jp/suisansc/suion.html; Air temperature data came

United States of America

Table 6.5: Preliminary predictions of V. parahaemolyticus illness in the United States of

6.1.3.8 Limitations and Caveats

6.1.4 Vibrio parahaemolyticus in bloody clams

O3:K6 tdh+, trh-

294/11 375 (2.6%)

The laboratory analysis undertaken included V. parahaemolyticus toxR gene sequencing to

To improve the risk assessment, the following data will be needed.

6.1.5 Vibrio parahaemolyticus in finfish

6.1.5.3 Hazard identification